Methods and compositions for prenatal testing

Abstract

The present disclosure provides methods and systems directed to isolation of circulating fetal cells (CFCs) from maternal biological samples of pregnancy-related states. A method for identifying genetic disorders of the fetus (or fetuses) of a subject may involve assaying a biological sample derived from the subject to isolate CFCs, and characterizing the biomarkers, such as the sequence of genetic material or abundance of gene expression material, from the CFCs to determine the presence or absence of a pregnancy-related states, such as a genetic order or ongoing health risk to the mother or fetus.

Description

WSGR Docket No. 62193-701.601 METHODS AND COMPOSITIONS FOR PRENATAL TESTING CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No.63 / 505,228, filed on May 31, 2023, which is incorporated by reference herein in its entirety for all purposes. BACKGROUND

[0002] Every year, about 7.9 million infants (e.g., 6% of worldwide births) are born with serious birth defect. Of those 7.9 million infants, about 3.2 million of these infants are disabled for life. Current prenatal testing methods for these birth defects are limited to imperfect (e.g., damaged) testing material (e.g., cell-free fetal DNA) and are unable to be performed until 10 weeks of pregnancy. SUMMARY

[0003] In some embodiments, the present disclosure provides fetal-specific biomarkers and maternal-specific biomarkers. In some embodiments, the fetal-specific biomarkers and the maternal-specific biomarkers can be used individually or in combinations to distingu ish circulating fetal cells from maternal cells in maternal blood samples. In some embodiments, the fetal-specific biomarkers and the maternal-specific biomarkers can purify the fetal cells from the maternal blood sample. In some embodiments, the purified fetal cells are used for prenatal genetic and gene expression analysis. In some embodiments, the present disclosure provides methods of purifying the circulating fetal cells from a maternal blood sample using the fetal- specific biomarkers and the maternal-specific biomarkers disclosed herein. In some embodiments, the circulating fetal cells can be purified from a maternal blood sample acquired during the first trimester of gestation. In some embodiments, the present disclosure provides methods for performing genetic analyses on the purified fetal cells. In some embodiments, the genetic analysis comprises fetal gene expression analysis. In some embodiments, the genetic analysis of the purified fetal cells can identify fetal genetic risk factors or gene expression markers of fetal health defects or pregnancy health defects.

[0004] In some embodiments, the present disclosure provides a composition comprising: an ex vivo population of cells from a maternal blood sample, wherein the ex vivo population of cells comprises a subpopulation of maternal cells and a subpopulation of fetal cells, wherein the subpopulation of fetal cells is at least about 75% of cells in the ex vivo population of cells.WSGR Docket No. 62193-701.601

[0005] In some embodiments, the subpopulation of fetal cells is at least about 80% of the cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 90% of the cells in the ex vivo population of cells.

[0006] In some embodiments, the subpopulation of maternal cells is no greater than about 30% of the cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 20% of the cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 10% of the cells in the ex vivo population of cells.

[0007] In some embodiments, the subpopulation of fetal cells comprises trophoblasts.

[0008] In some embodiments, the composition further comprises a second subpopulation of fetal cells, wherein the second subpopulation of fetal cells is different in cell type from the subpopulation of fetal cells. In some embodiments, the second subpopulation of fetal cells comprises erythroblasts. In some embodiments, the second subpopulation of fetal cells comprises megakaryocytes. In some embodiments, the second subpopulation of fetal cells comprises vascular endothelial cells. In some embodiments, the second subpopulation of fetal cells comprises fetal stromal cells. In some embodiments, the subpopulation of fetal cells comprises early gestation fetal cells, and the second subpopulation of fetal cells comprises later gestation fetal cells.

[0009] In some embodiments, the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by flow cytometry.

[0010] In some embodiments, the present disclosure provides a method comprising: sequencing a nucleic acid sequence, wherein the nucleic acid sequence is from an ex vivo population of cells from a maternal blood sample, wherein the ex vivo population of cells comprises a subpopulation of maternal cells and a subpopulation of fetal cells, wherein the subpopulation of fetal cells is at least about 70% of cells in the ex vivo population of cells.

[0011] In some embodiments, the subpopulation of fetal cells is at least about 80% of the cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 90% of the cells in the ex vivo population of cells.

[0012] In some embodiments, the subpopulation of maternal cells is no greater than about 30% of the cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 20% of the cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 10% of the cells in the ex vivo population of cells.WSGR Docket No. 62193-701.601

[0013] In some embodiments, the method further comprises, prior to the sequencing the nucleic acid sequence, extracting the subpopulation of maternal cells and the subpopulation of fetal cells from the maternal blood sample to provide an extract, wherein the extract comprises the subpopulation of maternal cells and the subpopulation of fetal cells from the maternal blood sample. In some embodiments, the method further comprises enriching the subpopulation of fetal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells. In some embodiments, the method further comprises sorting the subpopulation of fetal cells in the extract from the subpopulation of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

[0014] In some embodiments, prior to the sequencing the nucleic acid sequence, the subpopulation of maternal cells and the subpopulation of fetal cells are extracted from the maternal blood sample to provide an extract, wherein the extract comprises the subpopulation of maternal cells and the subpopulation of fetal cells from the maternal blood sample. In some embodiments, the subpopulation of fetal cells in the extract is enriched from the subpopulation of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells. In some embodiments, the subpopulation of fetal cells in the extract is sorted from the subpopulation of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

[0015] In some embodiments, the subpopulation of fetal cells comprises trophoblasts.

[0016] In some embodiments, the ex vivo population of cells further comprises a second subpopulation of fetal cells, wherein the second subpopulation of fetal cells is different in cell type from the subpopulation of fetal cells. In some embodiments, the second subpopulation of fetal cells comprises erythroblasts. In some embodiments, the second subpopulation of fetal cells comprises megakaryocytes. In some embodiments, the second subpopulation of fetal cells comprises vascular endothelial cells. In some embodiments, the second subpopulation of fetal cells comprises stromal cells. In some embodiments, the subpopulation of fetal cells comprises early gestation fetal cells and the second subpopulation of fetal cells comprises later gestation fetal cells.

[0017] In some embodiments, the method further comprises, prior to the sequencing the nucleic acid sequence, isolating the subpopulation of fetal cells from the maternal blood sample by contacting the maternal blood sample with a first isolation agent and coupling the f irst isolation agent to a first biomarker in the subpopulation of fetal cells. In some embodiments, the method further comprises incubating the maternal blood sample after the contacting. In some embodiments, the first isolation agent is a nucleic acid probe. In some embodiments, the firstWSGR Docket No. 62193-701.601 isolation agent is a ligand. In some embodiments, the first isolation agent couples to an intracellular biomarker. In some embodiments, the first isolation agent couples to a cell surface biomarker. In some embodiments, the first isolation agent couples to a soluble cell surface biomarker. In some embodiments, the first isolation agent couples to a nucleic acid biomarker.

[0018] In some embodiments, prior to the sequencing the nucleic acid sequence, the subpopulation of fetal cells is isolated from the maternal blood sample, wherein the maternal blood sample is contacted with a first isolation agent and the first isolation agent couples to a first biomarker in the subpopulation of fetal cells. In some embodiments, after the contacting, the maternal blood sample is incubated. In some embodiments, the first isolation agent is a nucleic acid probe. In some embodiments, the first isolation agent is a ligand. In some embodiments, the first isolation agent couples to a cell surface biomarker. In some embodiments, the first isolation agent couples to a soluble cell surface biomarker. In some embodiments, the first isolation agent couples to an intracellular biomarker. In some embodiments, the first isolation agent couples to a nucleic acid biomarker.

[0019] In some embodiments, prior to the sequencing the nucleic acid sequence, the subpopulation of maternal cells is isolated from the maternal blood sample, wherein the maternal blood sample is contacted with a second isolation agent and the second isolation agent couples to a second biomarker in the subpopulation of maternal cells. In some embodiments, after the contacting, the maternal blood sample is incubated. In some embodiments, the second isolation agent is a nucleic acid probe. In some embodiments, the second isolation agent is a ligand. In some embodiments, the second isolation agent couples to a cell surface biomarker. In some embodiments, the second isolation agent couples to a soluble cell surface biomarker. In some embodiments, the second isolation agent couples to an intracellular biomarker. In some embodiments, the second isolation agent couples to a nucleic acid biomarker.

[0020] In some embodiments, the method further comprises, prior to the sequencing the nucleic acid sequence, isolating the subpopulation of maternal cells from the maternal blood sample by contacting the maternal blood sample with a second isolation agent and coupling the second isolation agent to a second biomarker in the subpopulation of maternal cells. In some embodiments, the method further comprises incubating the maternal blood sample after the contacting. In some embodiments, the second isolation agent is a nucleic acid probe. In some embodiments, the second isolation agent is a ligand. In some embodiments, the second isolation agent couples to a cell surface biomarker. In some embodiments, the second isolation agent couples to a soluble cell surface biomarker. In some embodiments, the second isolation agentWSGR Docket No. 62193-701.601 couples to an intracellular biomarker. In some embodiments, the second isolation agent couples to a nucleic acid biomarker.

[0021] In some embodiments, the sequencing is performed on a next generation sequencing platform.

[0022] In some embodiments, the nucleic acid sequence is from the subpopulation of fetal cells. In some embodiments, the nucleic acid sequence is a DNA sequence. In some embodiments, the nucleic acid sequence is a RNA sequence.

[0023] In some embodiments, the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by flow cytometry.

[0024] In some embodiments, the present disclosure provides a method comprising: performing an assay on an ex vivo population of cells from a maternal blood sample, wherein the ex vivo population of cells comprises a subpopulation of maternal cells and a subpopulation of fetal cells, wherein the subpopulation of fetal cells is at least about 70% of cells in the ex vivo population of cells.

[0025] In some embodiments, the assay is a cell morphology assay. In some embodiments, the assay is a cell counting assay. In some embodiments, the assay is a cell viability assay. In some embodiments, the assay is a cell proliferation assay. In some embodiments, the assay is a cytotoxicity assay.

[0026] In some embodiments, the assay indicates a likelihood that a fetus associated with the subpopulation of fetal cells is susceptible to a genetic condition. In some embodiments, the genetic condition is down syndrome. In some embodiments, the genetic condition is cystic fibrosis. In some embodiments, the genetic condition is muscular dystrophy. In some embodiments, the genetic condition is sickle cell disease. In some embodiments, the genetic condition is Tay-Sachs disease.

[0027] In some embodiments, the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by flow cytometry. In some embodiments, the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by microscopy.

[0028] In some embodiments, the present disclosure provides a method comprising: (a) sequencing a first nucleic acid sequence obtained from a first type of fetal cell from an ex vivo population of fetal cells, (b) sequencing a second nucleic acid sequence obtained from a second type of fetal cell from the ex vivo population of fetal cells, (c) sequencing a third nucleic acid sequence obtained from a third type of fetal cell from the ex vivo population of fetal cells, (d) sequencing a fourth nucleic acid sequence obtained from a fourth type of fetal cell from the exWSGR Docket No. 62193-701.601 vivo population of fetal cells, (e) sequencing a fifth nucleic acid sequence obtained from a fifth type of fetal cell from the ex vivo population of fetal cells, and (f) analyzing at least one of the first nucleic acid sequence, the second nucleic acid sequence, the third nucleic acid sequence, the fourth nucleic acid sequence, or the fifth nucleic acid sequence to determine a likelihood that a fetus associated with an ex vivo population of fetal cells is susceptible to a genetic condition.

[0029] In some embodiments, at least one of the first nucleic acid sequence, the second nucleic acid sequence, the third nucleic acid sequence, the fourth nucleic acid sequence, or the fifth nucleic acid sequence comprises a DNA sequence. In some embodiments, at least one of the first nucleic acid sequence, the second nucleic acid sequence, the third nucleic acid sequence, the fourth nucleic acid sequence, or the fifth nucleic acid sequence comprises a RNA sequence.

[0030] In some embodiments, the ex vivo population of fetal cells is in a solution comprising a Artiodactyla serum.

[0031] In some embodiments, the method further comprises, prior to (a)-(f), extracting an ex vivo population of maternal cells and the ex vivo population of fetal cells from a maternal blood sample to provide an extract, wherein the extract comprises the ex vivo population of maternal cells and the ex vivo population of fetal cells. In some embodiments, the maternal blood sample is from a pregnant subject. In some embodiments, the maternal blood sample is blood plasma. In some embodiments, the maternal blood sample is blood serum. In some embodiments, the maternal blood sample is buffy coat. In some embodiments, the maternal blood sample comprises peripheral blood mononuclear white blood cells. In some embodiments, the maternal blood sample comprises nucleated white blood cells. In some embodiments, the maternal blood sample is whole blood. In some embodiments, the method further comprises enriching the ex vivo population of fetal cells in the extract from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells. In some embodiments, the method further comprises sorting the ex vivo population of fetal cells in the extract from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

[0032] In some embodiments, prior to (a)-(f), the ex vivo population of fetal cells and an ex vivo population of maternal cells are extracted from a maternal blood sample to provide an extract, wherein the extract comprises the ex vivo population of fetal cells and an ex vivo population of maternal cells. In some embodiments, the maternal blood sample is from a pregnant subject. In some embodiments, the maternal blood sample is blood plasma. In some embodiments, the maternal blood sample is blood serum. In some embodiments, the maternal blood sample is buffy coat. In some embodiments, the maternal blood sample comprises peripheral bloodWSGR Docket No. 62193-701.601 mononuclear white blood cells. In some embodiments, the maternal blood sample comprises nucleated white blood cells. In some embodiments, the maternal blood sample is whole blood. In some embodiments, the ex vivo population of fetal cells in the extract is enriched from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells. In some embodiments, the ex vivo population of fetal cells in the extract is sorted from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

[0033] In some embodiments, the method further comprises, prior to (a)-(f), incubating a blood sample comprising the ex vivo population of fetal cells and an ex vivo population of maternal cells with a first isolation agent and coupling the first isolation agent to a first biomarker in the ex vivo population of fetal cells. In some embodiments, the method further comprises sorting the ex vivo population of fetal cells based on a presence of the first isolation agent that couples to the first biomarker in the ex vivo population of fetal cells. In some embodiments, the method further comprises sorting the ex vivo population of fetal cells based on an absence of a second isolation agent that couples to a second biomarker in the ex vivo population maternal cells. In some embodiments, the first isolation agent is a nucleic acid probe. In some embodiments, the first isolation agent is a ligand. In some embodiments, the first isolation agent couples to a cell surface biomarker. In some embodiments, the first isolation agent couples to a soluble cell surface biomarker. In some embodiments, the first isolation agent couples to an intracellular biomarker. In some embodiments, the first isolation agent couples to a nucleic acid biomarker.

[0034] In some embodiments, prior to (a)-(f), a blood sample comprising the ex vivo population of fetal cells and an ex vivo population of maternal cells is incubated with a first isolation agent that couples to a first biomarker in the ex vivo population of fetal cells. In some embodiments, the ex vivo population of fetal cells is sorted based on a presence of the first isolation agent that couples to the first biomarker in the ex vivo population of fetal cells. In some embodiments, the ex vivo population of fetal cells is sorted based on an absence of a second isolation agent that couples to a second biomarker in the ex vivo population maternal cells. In some embodiments, the first isolation agent is a nucleic acid probe. In some embodiments, the first isolation agent is a ligand. In some embodiments, the first isolation agent couples to a cell surface biomarker. In some embodiments, the first isolation agent couples to a soluble cell surface biomarker. In some embodiments, the first isolation agent couples to an intracellular biomarker. In some embodiments, the first isolation agent couples to a nucleic acid biomarker.

[0035] In some embodiments, the method further comprises, prior to (a)-(f), incubating a blood sample comprising the ex vivo population of fetal cells and an ex vivo population of maternalWSGR Docket No. 62193-701.601 cells with a second isolation agent, and coupling the second isolation agent to a second biomarker in the ex vivo population of maternal cells. In some embodiments, the method further comprises sorting the ex vivo population of maternal cells based on a presence of the second isolation agent that couples to the second biomarker in the ex vivo population of maternal cells. In some embodiments, the method further comprises sorting the ex vivo population of maternal cells based on an absence of a first isolation agent that couples to a first biomarker in the ex vivo population of fetal cells. In some embodiments, the second isolation agent couples to an intracellular biomarker. In some embodiments, the second isolation agent is a nucleic acid probe. In some embodiments, the second isolation agent is a ligand. In some embodiments, the second isolation agent couples to a cell surface biomarker. In some embodiments, the second isolation agent couples to a soluble cell surface biomarker. In some embodiments, the second isolation agent couples to a nucleic acid biomarker.

[0036] In some embodiments, prior to (a)-(f), a blood sample comprising the ex vivo population of fetal cells and an ex vivo population of maternal cells is incubated with a second isolation agent that couples to a second biomarker in the ex vivo population of maternal cells. In some embodiments, the ex vivo population of maternal cells is sorted based on a presence of the second isolation agent that couples to the second biomarker in the ex vivo population of maternal cells. In some embodiments, the ex vivo population of maternal cells is sorted based on an absence of a first isolation agent that couples to a first biomarker in the ex vivo population of fetal cells. In some embodiments, the second isolation agent is a nucleic acid probe. In some embodiments, the second isolation agent is a ligand. In some embodiments, the second isolation agent couples to a cell surface biomarker. In some embodiments, the second isolation agent couples to a soluble cell surface biomarker. In some embodiments, the second isolation agent couples to an intracellular biomarker. In some embodiments, the second isolation agent couples to a nucleic acid biomarker.

[0037] In some embodiments, the ex vivo population of fetal cells comprises early gestation trophoblasts. In some embodiments, the ex vivo population of fetal cells comprises later gestation trophoblasts. In some embodiments, the ex vivo population of fetal cells comprises erythroblasts. In some embodiments, the ex vivo population of fetal cells comprises megakaryocytes. In some embodiments, the ex vivo population of fetal cells comprises stromal cells. In some embodiments, the ex vivo population of fetal cells comprises endothelial cells. In some embodiments, the ex vivo population of fetal cells comprises lymphocytes. In some embodiments, the ex vivo population of fetal cells comprises nucleated red blood cells. In some embodiments, the ex vivo population of fetal cells comprises hematopoietic progenitor cells. InWSGR Docket No. 62193-701.601 some embodiments, the ex vivo population of fetal cells comprises mesenchymal progenitor cells.

[0038] In some embodiments, the present disclosure provides a method comprising: performing a single round of enrichment of a population of fetal cells from a population of maternal cells in a cell sample, wherein the single round of enrichment enriches the population of fetal cells from the cell sample to a level that provides a quantity of fetal nucleic acid that is sufficient for characterization as fetal nucleic acid rather than maternal nucleic acid.

[0039] In some embodiments, the single round of enrichment provides an enriched sample, wherein the level that provides the quantity of fetal nucleic acid that is sufficient for characterization as fetal nucleic acid rather than maternal nucleic acid is at least about 50% of fetal cells of the enriched sample. In some embodiments, the method further comprises performing one and no greater than one round of the enrichment.

[0040] In some embodiments, the cell sample is a blood sample. In some embodiments, the blood sample is blood plasma. In some embodiments, the blood sample is blood serum. In some embodiments, the blood sample is buffy coat. In some embodiments, the maternal blood sample comprises peripheral blood mononuclear white blood cells. In some embodiments, the maternal blood sample comprises nucleated white blood cells. In some embodiments, the blood sample is whole blood.

[0041] In some embodiments, the cell sample is from a human subject.

[0042] In some embodiments, the method further comprises, prior to the performing, extracting the population of maternal cells and the population of fetal cells from the cell sample to provide an extract, wherein the extract comprises the population of maternal cells and the population of fetal cells. In some embodiments, the method further comprises sorting the population of fetal cells in the extract from the population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

[0043] In some embodiments, prior to the performing, the population of fetal cells and the population of maternal cells are extracted from the cell sample to provide an extract, wherein the extract comprises the population of maternal cells and the population of fetal cells. In some embodiments, the population of fetal cells in the extract is sorted from the population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

[0044] In some embodiments, prior to the performing, the cell sample is incubated with a first isolation agent specific for a first biomarker in the population of feta l cells. In some embodiments, the first isolation agent is a nucleic acid probe. In some embodiments, the firstWSGR Docket No. 62193-701.601 isolation agent is a ligand. In some embodiments, the first biomarker comprises a cell surface marker. In some embodiments, the first biomarker comprises a soluble cell surface marker. In some embodiments, the first biomarker comprises an intracellular biomarker. In some embodiments, the first biomarker comprises a nucleic acid marker.

[0045] In some embodiments, the method further comprises, prior to the performing, incubating the cell sample with a first isolation agent specific for a first biomarker in the population of fetal cells. In some embodiments, the first isolation agent is a nucleic acid probe. In some embodiments, the first isolation agent is a ligand. In some embodiments, the first biomarker comprises a cell surface marker. In some embodiments, the first biomarker comprises a soluble cell surface marker. In some embodiments, the first biomarker comprises an intracellular biomarker. In some embodiments, the first biomarker comprises a nucleic acid marker.

[0046] In some embodiments, prior to the performing, the cell sample is incubated with a second isolation agent specific for a second biomarker in the population of maternal cells. In some embodiments, the second isolation agent is a nucleic acid probe. In some embodiments, the second isolation agent is a ligand. In some embodiments, the second biomarker comprises a cell surface marker. In some embodiments, the second biomarker comprises a soluble cell surface marker. In some embodiments, the second biomarker comprises an intracellular biomarker. In some embodiments, the second biomarker comprises a nucleic acid marker.

[0047] In some embodiments, the method further comprises, prior to the performing, incubating the cell sample with a second isolation agent specific for a second biomarker in the population of maternal cells. In some embodiments, the second isolation agent is a nucleic acid probe. In some embodiments, the second isolation agent is a ligand. In some embodiments, the second biomarker comprises a cell surface marker. In some embodiments, the second biomarker comprises a soluble cell surface marker. In some embodiments, the second biomarker comprises an intracellular biomarker. In some embodiments, the second biomarker comprises a nucleic acid marker.

[0048] In some embodiments, the method further comprises sequencing a nucleic acid sequence from the population of fetal cells.

[0049] In some embodiments, the present disclosure provides a method comprising: a) extracting an ex vivo subpopulation of maternal cells and an ex vivo subpopulation of fetal cells from a maternal blood sample, b) after the extracting, enriching the ex vivo subpopulation of fetal cells from the ex vivo subpopulation of maternal cells, c) after the enriching, sorting the ex vivo subpopulation of fetal cells from the ex vivo subpopulation from maternal cells to provide aWSGR Docket No. 62193-701.601 sample enriched in fetal cells and depleted in maternal cells, and d) after the sorting, sequencing a nucleic acid from the ex vivo subpopulation of fetal cells.

[0050] In some embodiments, the present disclosure provides a method comprising: a) contacting a maternal blood sample comprising an ex vivo subpopulation of fetal cells and an ex vivo subpopulation of maternal cells with an isolation agent that couples to a biomarker in the ex vivo subpopulation of fetal cells, b) after the contacting, incubating the isolation agent with the maternal blood sample, c) after the incubating, isolating the ex vivo subpopulation of fetal cells in the maternal blood sample from the ex vivo subpopulation of maternal cells in the maternal blood sample, and d) after the isolating, sequencing a nucleic acid from the ex vivo subpopulation of fetal cells.

[0051] In some embodiments, the present disclosure provides a method comprising: a) contacting a maternal blood sample comprising an ex vivo subpopulation of fetal cells and an ex vivo subpopulation of maternal cells with an isolation agent that couples to a biomarker in the ex vivo subpopulation of fetal cells, b) after the contacting, incubating the isolation agent with the maternal blood sample, c) after the incubating, isolating the ex vivo subpopulation of fetal cells in the maternal blood sample from the ex vivo subpopulation of maternal cells in the maternal blood sample, d) after the isolating, sorting the ex vivo subpopulation of fetal cells from the ex vivo subpopulation of maternal cells, and e) after the sorting, sequencing a nucleic acid from the ex vivo subpopulation of fetal cells.

[0052] In an aspect, the present disclosure provides a method comprising: incubating a population of cells from a sample of a subject with a first isolation agent, a second isolation agent, a third isolation agent, a fourth isolation agent, a fifth isolation agent, and a sixth isolation agent, wherein the population of cells comprises a subpopulation of fetal cells and a subpopulation of maternal cells, wherein the first isolation agent is specific for a first biomarker, the second isolation agent is specific for a second biomarker, the third isolation agent is specific for a third biomarker, the fourth isolation agent is specific for a fourth biomarker, and the fifth isolation agent is specific for a fifth biomarker in the subpopulation of fetal cells, and wherein the sixth isolation agent is specific for a sixth biomarker in the subpopulation of maternal cells.

[0053] In some embodiments, the subject is a human subject.

[0054] In some embodiments, the sample is a blood sample. In some embodiments, the blood sample is blood plasma. In some embodiments, the blood sample is blood serum. In some embodiments, the blood sample is buffy coat. In some embodiments, the maternal blood sample comprises peripheral blood mononuclear white blood cells. In some embodiments, the maternalWSGR Docket No. 62193-701.601 blood sample comprises nucleated white blood cells. In some embodiments, the blood sample is whole blood.

[0055] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is a nucleic acid probe.

[0056] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is a ligand.

[0057] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a cell surface marker.

[0058] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a soluble cell surface marker.

[0059] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for an intracellular biomarker.

[0060] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the s ixth isolation agent is specific for a nucleic acid marker.

[0061] In some embodiments, the method further comprises, after the incubating, enriching the subpopulation of fetal cells from the subpopulation of maternal cells.

[0062] In some embodiments, after the incubating, the subpopulation of fetal cells is enriched from the subpopulation of maternal cells.

[0063] In some embodiments, the present disclosure provides a method comprising: incubating a population of cells a first isolation agent, a second isolation agent, a third isolation agent, a fourth isolation agent, a fifth isolation agent, and a sixth isolation agent, wherein the population of cells comprises a subpopulation of fetal cells and a subpopulation of maternal cells from a sample of a subject, and wherein the first isolation agent is specific for a first biomarker in the subpopulation of fetal cells, the second isolation agent is specific for a second biomarker in the subpopulation of fetal cells, the third isolation agent is specific for a third biomarker in the subpopulation of fetal cells, the fourth isolation agent is specific for a fourth biomarker in the subpopulation of maternal cells, the fifth isolation agent is specific for a fifth biomarker in theWSGR Docket No. 62193-701.601 subpopulation of maternal cells, and the sixth isolation agent is specific for a sixth biomarker in the subpopulation of maternal cells.

[0064] In some embodiments, the subject is a human subject.

[0065] In some embodiments, the sample is a blood sample. In some embodiments, the blood sample is blood plasma. In some embodiments, the blood sample is blood serum. In some embodiments, the blood sample is buffy coat. In some embodiments, the maternal blood sample comprises peripheral blood mononuclear white blood cells. In some embodiments, the maternal blood sample comprises nucleated white blood cells. In some embodiments, the blood sample is whole blood.

[0066] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the six th isolation agent is a nucleic acid probe.

[0067] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is a ligand.

[0068] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a cell surface marker.

[0069] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a soluble cell surface marker.

[0070] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for an intracellular biomarker.

[0071] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a nucleic acid marker.

[0072] In some embodiments, the method further comprises, after the incubating, enriching the subpopulation of fetal cells from the subpopulation of maternal cells.

[0073] In some embodiments, after the incubating, the subpopulation of fetal cells is enriched from the subpopulation of maternal cells.

[0074] In some embodiments, the present disclosure provides a method comprising: incubating a population of cells with a first isolation agent, a second isolation agent, a third isolation agent, a fourth isolation agent, a fifth isolation agent, a sixth isolation agent, and a seventh isolationWSGR Docket No. 62193-701.601 agent, wherein the population of cells comprises a subpopulation of fetal cells and a subpopulation of maternal cells from a sample of a subject, wherein the first isolation agent is specific for a first biomarker, the second isolation agent is specific for a second biomarker, the third isolation agent is specific for a third biomarker, the fourth isolation agent is specific for a fourth biomarker, the fifth isolation agent is specific for a fifth biomarker, and the sixth isolation agent is specific for a sixth biomarker in the subpopulation of maternal cells, and wherein the seventh isolation agent is specific for a seventh biomarker in the subpopulation of fetal cells.

[0075] In some embodiments, the subject is a human subject.

[0076] In some embodiments, the sample is a blood sample. In some embodiments, the blood sample is blood plasma. In some embodiments, the blood sample is blood serum. In some embodiments, the blood sample is buffy coat. In some embodiments, the maternal blood sample comprises peripheral blood mononuclear white blood cells. In some embodiments, the maternal blood sample comprises nucleated white blood cells. In some embodiments, the blood sample is whole blood.

[0077] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent comprises a nucleic acid probe.

[0078] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent comprises a ligand.

[0079] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent is specific for a cell surface marker.

[0080] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent is specific for a soluble cell surface marker.

[0081] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation, the sixth isolation agent, and the seventh isolation agent is specific for an intracellular biomarker.

[0082] In some embodiments, one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent is specific for a nucleic acid marker.

[0083] In some embodiments, the method further comprises, after the incubating, enriching the subpopulation of fetal cells from the subpopulation of maternal cells.WSGR Docket No. 62193-701.601

[0084] In some embodiments, after the incubating, the subpopulation of fetal cells is enriched from the subpopulation of maternal cells.

[0085] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in this art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive. INCORPORATION BY REFERENCE

[0086] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material. BRIEF DESCRIPTION OF THE DRAWINGS

[0087] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0088] FIG. 1 illustrates a schematic for mouse breeding and experimental protocol for the identification of fetal cell-specific biomarkers and maternal cell-specific biomarkers.

[0089] FIG.2 illustrates flow plots depicting the isolation of fetal cells from a maternal blood sample. The 201 plot depicts the total number of cells in the maternal blood sample. The 202 plot depicts the total number of single cells (e.g., not clustered cells) of the total number of cells in the maternal blood sample. The 203 plot depicts the number of viable cells (e.g., Calcein violet-positive cells) of the single cells in the maternal blood sample. The 204 plot depicts the number of nucleated cells (e.g., DyeCycle-positive cells) in the viable cells in the maternal blood sample. The 205 plot depicts the number of live cells (e.g., Sytox-orange-negative cells) in the nucleated cells in the maternal blood sample. The 206 table shows the number of cells in each of 201, 202, 203, 204, 205, and 207 plots. The 207 plot shows the number of circulating fetal cells (e.g., eGFP-positive cells) I the live cells in the maternal blood sample.WSGR Docket No. 62193-701.601

[0090] FIG. 3A depicts a single-cell gene expression analysis of maternal blood enriched for eGFP-expressing fetal cells. The clusters of cells enclosed by the black circles are the cells with the most robust expression of fetal-specific biomarkers.

[0091] FIG. 3B depicts single-cell gene expression profiles. The cells enclosed in the black circles are determined to be fetal trophoblasts, fetal stromal cells, and additional fetal cell types by single-cell gene expression analysis. The clusters of cells not enclosed in the black circles are determined to be different maternal cell types (e.g., megakaryocytes, B cells, T cells, NK cells, Basophils, Dendritic cells, macrophages, neutrophils, and erythroblasts).

[0092] FIG.4A depicts the identification of fetal cells using a combination of fetal cell-specific biomarkers (left panel) against a background of maternal blood cell types. The right panel depicts the identification of different fetal cell types using a combination of fetal cell-specific biomarkers and maternal cell-specific biomarkers against a background of maternal blood cell types. FIG.4B depicts the detection of early gestation stromal cells using a combination of fetal cell-specific biomarkers and maternal cell-specific biomarkers (left panel). The right panel depicts the identification of early gestation trophoblasts and late gestation trophoblasts using fetal cell-specific biomarkers.

[0093] FIG. 5 depicts a high-resolution clustering model of the isolated CFCs. The clustering model shows that placenta-related CFCs and hematopoietic CFCs are distinguished from each other based on different expression ratios of the fetal cell-specific cell biomarker Malat1.

[0094] FIG. 6A-6H depict heatmaps showing the patterns of cell type-specific expression of fetal cell-specific biomarkers and maternal cell-specific biomarkers. The x-axis of the heatmap depicts the different cell types (e.g., B cells, erythroblasts, macrophages, megakaryocytes, neutrophils, stromal cells, T cells, trophoblasts) and y-axis shows the name of the fetal cell- specific biomarker or the maternal cell-specific biomarker.

[0095] FIG. 7A depicts the purity and yield of circulating fetal cells (CFCs) isolated from a maternal blood sample using sequential positive selections for fetal cells that are double-positive for two fetal biomarkers.

[0096] FIG. 7B depicts the purity and yield of CFCs isolated from a maternal blood sample using sequential positive selections for fetal cells that are double-positive for two fetal biomarkers.

[0097] FIG. 7C depicts the purity and yield of CFCs isolated from a maternal blood sample using sequential positive selections from fetal cells using two different combinations of fetal biomarkers.WSGR Docket No. 62193-701.601

[0098] FIG. 7D depicts the purity and yield of CFCs isolated from a maternal blood sample using sequential positive selections from fetal cells using two different combinations of fetal biomarkers.

[0099] FIG. 7E depicts the purity and yield of CFCs isolated from a maternal blood sample using a combination of negative selection with a combination of maternal biomarkers and sequential positive selections using two different combinations of fetal biomarkers. DETAILED DESCRIPTION

[0100] While various embodiments of the invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed.

[0101] In some embodiments, the present disclosure provides methods, compositions, and kits for isolating fetal cells from a maternal sample. In some embodiments, fetal cells and maternal cells are isolated from a maternal sample. In some embodiments, the fetal cells are enriched (e.g., sorted) from the maternal cells. In some embodiments, the fetal cells are enriched using isolation agents. In some embodiments, the fetal cells are incubated (e.g., contacted) with the isolation agents. In some embodiments, the isolation agents couple to a fetal cell-specific biomarker. In some embodiments, fetal nucleic acid molecules are extracted from the fetal cells. In some embodiments, the fetal nucleic acid molecules are sequenced. In some embodiments, the sequenced fetal nucleic acid molecules are used for prenatal testing. In some embodiments, the prenatal testing determines the presence or absence of genetic disorders or conditions. In some embodiments, the fetal cells are used to perform an assay. Compositions Cellular Compositions

[0102] In some embodiments, the present disclosure provides an ex vivo population of cells from a subject. In some embodiments, the ex vivo population of cells from the subject is from a pregnant subject. In some embodiments, the pregnant subject has two types of cells. In some embodiments, the two types of cells are fetal cells and maternal cells. In some embodiments, the fetal cells are cells that are derived from the fetus and / or the placenta. In some embodiments, the fetal cells have genetic information from the mother and genetic information from the father. In some embodiments, the maternal cells are cells that are derived from maternal organs and tissues. In some embodiments, the maternal cells have genetic information from the mother. In some embodiments, the maternal cells do not have genetic information from the father. In someWSGR Docket No. 62193-701.601 embodiments, a sample is taken from the pregnant subject. In some cases, the sample has fetal cells and maternal cells. In some cases, the ex vivo population of cells is isolated from the sample taken from the pregnant subject.

[0103] In some embodiments, the ex vivo population of cells is isolated from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a mouse. In some embodiments, the subject is a rat. In some embodiments, the subject is a non -human primate. In some embodiments, the subject is a human. In some embodiments, the subject is pregnant. In some embodiments, the sample is a biological sample. In some embodiments, the biological sample is a blood sample. In some embodiments, the biological sample is a tissue and / or biopsy sample. In some embodiments, the tissue and / or biopsy sample is from the uterus, placenta, umbilical cord, or any combination thereof. In some embodiments, the biological sample is a biological fluid sample. In some embodiments, the biological fluid sample is blood, saliva, interstitial fluid, umbilical cord blood, extracellular fluid, or any combination thereof .

[0104] In some embodiments, the ex vivo population of cells is isolated through density gradient centrifugation. In some embodiments, the ex vivo population of cells is isolated through density sedimentation. In some embodiments, the ex vivo population of cells is isolated through immunomagnetic cell separation. In some embodiments, the ex vivo population of cells is isolated through fluorescence-activated cell sorting (FACS). In some embodiments, the ex vivo population of cells is isolated through magnetic-activated cell sorting (MACS). In some embodiments, the ex vivo population of cells is isolated through microscopic microdissection. In some embodiments, the ex vivo population of cells is isolated through microscopic imaging. In some embodiments, the ex vivo population of cells is isolated from plasma and / or platelets from the blood sample.

[0105] In some embodiments, the ex vivo population of cells has a subpopulation of maternal cells. Non-limiting examples of maternal cell include red blood cells, T cells, B cells, dendritic cells, macrophages, monocytes, erythrocytes, neutrophils, and lymphocytes. In some embodiments, the ex vivo population of cells has a subpopulation of fetal cells. In some embodiments, the subpopulation of fetal cells is circulating fetal cells (CFCs). CFCs can be fetal cells that circulate in the maternal bloodstream. In some embodiments, the subpopulation of fetal cells contains one or more subpopulations of fetal cells, for example, a first subpopulation and a second subpopulation. In some embodiments, the second subpopulation of fetal cells is different in cell type from the first subpopulation of fetal cells. In some embodiments, the first subpopulation of fetal cells comprises early gestation fetal cells and the second subpopulation of fetal cells comprises later gestation fetal cells. Non-limiting examples of types of fetal cellsWSGR Docket No. 62193-701.601 include early gestation trophoblasts (e.g., early gestation trophoblasts), later gestation trophoblasts (e.g., late gestation trophoblasts), erythroblasts, megakaryocytes, stromal cells, endothelial cells, lymphocytes, nucleated red blood cells, hematopoietic progenitor cells, vascular endothelial cells, and mesenchymal progenitor cells.

[0106] In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises later gestation trophoblasts. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises erythroblasts. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises erythroblasts. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises megakaryocytes. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises megakaryocytes. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises stromal cells. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises stromal cells. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises endothelial cells. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises endothelial cells. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises lymphocytes. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises lymphocytes. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In someWSGR Docket No. 62193-701.601 embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises early gestation trophoblasts and the second subpopulation of fetal cells comprises vascular endothelial cells. In some embodiments, the first subpopulation of fetal cells comprises later gestation trophoblasts and the second subpopulation of fetal cells comprises vascular endothelial cells.

[0107] In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises megakaryocytes. In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises stromal cells. In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises endothelial cells. In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises lymphocytes. In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises erythroblasts and the second subpopulation of fetal cells comprises vascular endothelial cells.

[0108] In some embodiments, the first subpopulation of fetal cells comprises megakaryocytes and the second population of fetal cells comprises stromal cells. In some embodiments, the first subpopulation of fetal cells comprises megakaryocytes and the second population of fetal cells comprises endothelial cells. In some embodiments, the first subpopulation of fetal cells comprises megakaryocytes and the second population of fetal cells comprises lymphocytes. In some embodiments, the first subpopulation of fetal cells comprises megakaryocytes and the second subpopulation of fetal cells comprises nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells comprises megakaryocytes and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises megakaryocytes and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises megakaryocytes and the second subpopulation of fetal cells comprises vascular endothelial cells.WSGR Docket No. 62193-701.601

[0109] In some embodiments, the first subpopulation of fetal cells comprises stromal cells and the second subpopulation of fetal cells comprises endothelial cells. In some embodiments, the first subpopulation of fetal cells comprises stromal cells and the second subpopulation of fetal cells comprises lymphocytes. In some embodiments, the first subpopulation of fetal cells comprises stromal cells and the second subpopulation of fetal cells comprises nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells comprises stromal cells and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises stromal cells and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises stromal cells and the second subpopulation of fetal cells comprises vascular endothelial cells.

[0110] In some embodiments, the first subpopulation of fetal cells comprises endothelial cells and the second subpopulation of fetal cells comprises lymphocytes. In some embodiments, the first subpopulation of fetal cells comprises endothelial cells and the second subpopulation of fetal cells comprises nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells comprises endothelial cells and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises endothelial cells and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises endothelial cells and the second subpopulation of fetal cells comprises vascular endothelial cells.

[0111] In some embodiments, the first subpopulation of fetal cells comprises lymphocytes and the second subpopulation of fetal cells comprises nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells comprises lymphocytes and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises lymphocytes and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises lymphocytes and the second subpopulation of fetal cells comprises vascular endothelial cells.

[0112] In some embodiments, the first subpopulation of fetal cells comprises nucleated red blood cells and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises nucleated red blood cells and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises nucleated red blood cells and the second subpopulation of fetal cells comprises vascular endothelial cells.WSGR Docket No. 62193-701.601

[0113] In some embodiments, the first subpopulation of fetal cells comprises hematopoietic progenitor cells and the second subpopulation of fetal cells comprises mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises mesenchymal progenitor cells and the second subpopulation of fetal cells comprises hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells comprises mesenchymal progenitor cells and the second subpopulation of fetal cells comprises vascular endothelial cells. In some embodiments, the first subpopulation of fetal cells comprises hematopoietic progenitor cells and the second subpopulation of fetal cells comprises vascular endothelial cells. In some embodiments, the ex vivo population of cells has subpopulation of fetal cells and a subpopulation of maternal cells.

[0114] In some embodiments, the subpopulation of fetal cells is from about 50% to about 100% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is from about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, or more of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at most about 100%, at most about 95%, at most about 90%, at most about 85%, at most about 80%, at most about 75%, at most about 70%, at most about 65%, at most about 60%, at most about 55%, at most about 50%, or less of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 50% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 60% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 70% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 80% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 90% of cells in the ex vivo population of cells.

[0115] In some embodiments, the subpopulation of maternal cells is no greater than about 0%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 50% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 40% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 30% of cells in the ex vivo population of cells. In someWSGR Docket No. 62193-701.601 embodiments, the subpopulation of maternal cells is no greater than about 20% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 10% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 5% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of maternal cells is no greater than about 0% of cells in the ex vivo population of cells.

[0116] In some embodiments, the ex vivo population of cells has a purity between about 50% to about 100% of fetal cells. In some embodiments, the ex vivo population of cells has a purity from about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of fetal cells. In some embodiments, the ex vivo population of cells has a purity of at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, or more of fetal cells. In some embodiments, the ex vivo population of cells has a purity of at most about 100%, at most about 95%, at most about 90%, at most about 85%, at most about 80%, at most about 75%, at most about 70%, at most about 65%, at most about 60%, at most about 55%, at most about 50%, or less of fetal cells. In some embodiments, the ex vivo population of cells has a purity from about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of fetal cells.

[0117] In some embodiments, the subpopulation of fetal cells has one or more fetal cell biomarkers. In some embodiments, the one or more biomarkers for the subpopulation of fetal cells are different from one or more maternal cell biomarkers. Non-limiting examples of fetal cell biomarkers include H19, 1600025M17Rik, Sct, Ctsr, Rhox9, Prl2c5, Rhox6, Fthl17a, Ctsj, Prl3b1, Nrk, Prl2b1, Ctsq, Hand1, Rhox5, Prl2c2, Krt8, Prl2c3, Prl7a1, Trap1a, Prl3d1, Igf2, Krt18, Nup62cl, Prl7d1, Rhox12, Prl2a1, Krt19, Plac1, Prl3d2, Serpinb9g, Slc38a4, Prl4a1, Serpinb9d, Serpinb9e, Prl8a9, Cited1, Sparc, Gm9112, Ldoc1, Serpinb9f, 3830417A13Rik, Serpinb9c, Tinagl1, Tfpi, Dusp9, Cts6, Hspb1, Nxf7, Sult1e1, Fnd3c2, Rarres2, Procr, Fermt2, Ghrh, Prl2c1, Plet1, Sbsn, Cdh5, Psg19, Igf2bp1, Tpbpa, Cda, Erv3, Pramef12, Shroom1, Peg10, 1700011M02Rik, Psg29, Cyr61, Crct1, Prl8a1, Psg22, Chrdl2, Xlr5a, Prl3d3, Ceacam9, Inhbb, Krt7, Serpinb9b, Cdkn1c, Rbp1, Mrgprg, Gm7120, Aoc1, Pla2g5, Rhox1, Tfap2c, A2m, Tpbpb, Lmcd1, Cryaa, Prl5a1, Zfp36l3, Gm9, Prl6a1, Rgs17, Bex1, Cdh3, Prl7b1, Hsd17b2, Cyp17a1, Gm9513, Tex19.1, Parva, Papl, Map1b, Gpr50, 1600015l10Rik, Ceacam13, Prom2, Slco5a1, Cts7, Nos2, Nppc, Arhgap42, Ppp2r2c, Peg3, Mfap5, Tnfrsf9, Prl8a2, Ceacam15, Rimklb, Aard, Prl7a2, Slc30a2, 4933402E13Rik, Mmp1a, Epn3, Irx3, Fam83b, Tead4, Lama1,WSGR Docket No. 62193-701.601 Htra1, Pfpl, Cts8, Adgrf5, Prl8a6, Drd4, Bmp1, Cystm1, Gml2, Gm13889, Krtdap, Pappa2, Sema3e, Homer2, Krt14, Pla2g2f, Klhl13, 2310030G06Rik, Car4, Gm7257, Prss8, Dsc2, Ptprn2, Des, Hsd3b6, Pxdn, 270033N17Rik, Eps8l2, 1700001C19Rik, Ceacam11, 1700089L19Rik, Trpm5, 1600012P17Rik, Got1l1, Dmrtc1b, Mt4, Spint1, Mt2, Fscn1, Gm14547, Serpine1, Col4a1, Ndufa4l2, Dst, Pvrl2, Doxl2, Cyp11a1, Klk6, Atf7ip2, Gm805, Pou2f3, Gpr173, Afap1l2, Nkain1, Fabp6, Triml2, Cryab, Pdgfra, Psca, Scnn1b, Rab15, Fmr1nb, Adamts7, Igfbpl1, A4galt, Hrct1, Aif1l, Gm16136, Sema3f, Fam189a1, Psg27, Gm22, Gm26829, Igf2os, Rpl29, Eif2s3y, Il25, Actn3, Sez6l, Unc5d, Frem2, Gm20714, Tff1, Hrc, Hspg2, Pdia5, Ccdc155, 2310016G11Rik, Fam90a1b, Vgf, Cts3, Klk4, Ttc9, Ces1a, Zfp385b, 4631405K08Rik, Nptx2, Adra2b, Mettl24, Gm10251, Gm4758, Prl7c1, Col4a2, Cmtm5, Il24, Ephb3, Cartpt, Foxf1, Gm11755, Spdye4a, Myt1l, 4930562A09Rik, 9930038B18Rik, Msx1, Elf3, Adgrf4, Dio3, Rgs5, Star, Bcat1, Podxl, Lepr, Col5a3, Slco2a1, Myom3, Gm9979, Gpnmb, Fam159a, Trim29, Aqp8, Syt9, Kng2, Wnt7b, St8sia5, Tmem125, Dysf, Uaca, Fetub, Bcar1, Pcdh12, Gm14393, Fabp3, Stra6, Osmr, Lamc2, Muc1, RP23-378L12.3, Bok, Fdx1, Pkp2, Acbd7, Cyb5r3, Sdc1, Sfrp5, Lama5, Slc39a2, Zfp42, Sox3, Vvstm4, Gpx3, Dmkn, Gjb3, Lgi2, Stard10, Slc6a2, Prune2, H19, Sct, Rhox9, Krt18, Prl2c5, Rhox6, Krt8, Prl2c3, Ctsj, Krt19, Prl3b1, Prl3d1, Prl7d1, Prl7a1, Tfpi, Trap1a, Fthl17a, Ctsr, Sparc, Igf2, Prl2c2, Hspb1, Nrk, Ctsq, Rhox5, Prl2b1, Rarres2, Hand1, Peg10, Prl2a1, Procr, Sbsn, Prl4a1, Rhox12, Cdkn1c, Ghrh, Cdh5, Cited1, Gm9112, Krt7, Plac1, 1600025M17Rik, Serpinb9d, Cts6, Prl3d2, Serpinb9e, Dusp9, Map1b, Plet1, Nup62cl, Serpinb9g, Fermt2, Slc38a4, Ldoc1, Serpinb9f, 3830417A13Rik, Prl8a1, Hsd17b2, Prl8a9, Psg29, Nxf7, Psg22, Fnd3c2, Serpinb9c, Bex1, 1700011M02Rik, Tpbpa, Prl3d3, Prl2c1, Parva, Tinagl1, Prl5a1, Adgrf5, Igf2bp1, Rbp1, Mt2, Serpinh1, Tfap2c, Serpinb9b, Klhl13, Prl8a2, Sult1e1, Cyp11a1, Gjb3, Gm7120, Htra1, Cyp17a1, Cystm1, Pramef12, Tnfrsf9, Hsd3b6, Crip2, Ceacam9, Lgals1, Ctsl, Cryab, Homer2, Slc30a2, Cdh3, Zfp36l3, Nppc, Ceacam15, Syngr1, Shroom1, Fscn1, Hspg2, Fmr1nb, Mmp1a, Sdc1, Pfpl, Krt14, Tex19.1, Psg19, Cb5r3, Sema3e, Peg3, Pdia5, Chrdl2, Dst, Dbi, Meis2, Sema3f, Rgs17, Col4a2, Fdx1, Dmrtc1b, Slco2a1, Tpm1, Stard10, Mdk, Irx3, Maged1, Star, Csrp1, Nbl1, Rnd2, Prdx4, Des, Alpl, Xlr5a, Afao1l2, Aprt, Arhgap42, Vgf, Erv3, Tbpbp, Cst8, P4hb, Tbrg1, Cyr61, Muc1, Tmem37, Podxl, Fabp3, Fkbp9, Fstl3, Car4, Epas1, Tpm2, Rhox1, Chchd10, Stra6, Egfl7, Rab15, Copz2, Nos2, Lgals3, Sin3b, Slc6a2, Epcam, Pkp2, Pla2g5, Pvrl2, Rhox10, Prl7b1, Ada, Lmcd1, Erdr1, Selm, 1700086L19Rik, Dmrtc1a, Bok, Ldhb, Epn3, Mrgprg, Basp1, Gm9, Aoc1, Serpinb6b, Uaca, 2310030G06Rik, Ckb, Cstb, Ssr4, Foxo4, Ngef, Plpp1, A2m, Dstn, Cox6a1, Gale, Pde10a, Krtdap, Rab6b, Esam, Crct1, Col5a2, A4galt, Dnase1l3, Sfn, Bmp1, Calr, Papl, Kank1, Prl6a1, Cmtm5, Ildr2, Rhoc, Ccl27a, Efs, Hrct1, Dsc2,WSGR Docket No. 62193-701.601 Bcar1, Cd63, Inhbb, Ppp2r2c, Mpdz, Mt1, Fetub, Nid1, Bicc1, Psg23, Hdlbp, Rasd1, Arhgap22, Maged2, Aard, Prl8a6, Cald1, Prom2, Pdia6, Nostrin, Prss8, 1600012P17Rik, Dsp, Adm, Fam90a1b, Exoc3l4, Rbfox2, Mgst3, Gm648, Rgs5, Bgn, Galk1, Adra2b, AT036118, Nenf, Aif1l, Nfib, Acvr2b, Htatsf1, Rpn1, Psg27, Phldb2, Myof, Fbxl19, H1f0, Cobl, Emp2, Myrf, Adamts7, Fam89a, Gm9513, Tmem98, Mfsd2a, Mgat4b, Cav1, Il33, C77080, Lars2, Gm21887, 9930038B18Rik, Prom1, Ptov1, Ltbp4, Mmp23, Prl7a2, Cts3, Fabp6, Prkcdbp, Lmna, Rpn2, Caskin1, Lama1, Fam83b, Cryaa, Lamc2, 4933402E13Rik, Fn1, Dlc1, Lrrn4, Tff1, Elf3, Serpine1, Bcat1, Perp, Lama5, Gpx3, Car2, Pawr, Phactr1, Oaf, Lamb2, Prnp, Scin, Parm1, Smim1, Jup, Gpx8, Atf5, Ift43, Gkap1, H1fx, Cenpt, Rhou, Rrm2, Cul7, Asns, Tubb3, 2200002D01Rik, Abhd6, Hdac6, Rgs16, Cgnl1, Atp9a, Src, 4930486L24Rik, Elf3, Ly6c1, transferrin receptor (TfR), ZNF614, ZNF539, ZNF283, ZFYVE9, WRN, UBL4A, TPM3, TP73, TMEFF2, THC2274391, THC2265980, t (BRACHYURY), SYT9, SLAMF1, RPS27, RPL39, RPL26, RPL23A, RPL23, RP11-78J21.1, RGPD2, QPRT, PPIA, PGK1, PF4, NTRK1, NBR2, MYL6, MAPK3, MAD2L1, LOC389286, KY, KMO, KCNK4, ITGA7, HMGN2, HMGA1, HLA-C, HIST1H2AJ, GSK3A, GCC2, FYN, F1135740, ENST00000356196, EEF1A1, EDN1, DKFZP434F142, D4ST1, CRYL1, CPS1, CD7, CCNA2, CB123670, BC111482, 6C089454, ALS2CL, AK000420, AHNAK, ADAM11, ACVR2B, ABHD2, CD105, HLA-G, galectin 13 (LGAGLS13 / PP13), galectin 14 (LGALS14), placental growth factor (PGF), pregnancy- associated plasma protein A (PAPPA), alpha fetal protein (AFP), endoglin (ENG), fns-related tyrosine kinase 1 (FLT-1), keratin-7, PLAP, MCAM, laeverin, H315, FT1.41.1, NDOG-1, NDOG-5, BC1, AB-154, AB-340 (PAR-1), Glut-12, factor XIII, hPLH, HLA-C, JunD, Fra2, NDPK-A, CAR, HASH2, αHCG, IGF-11, PAI-1, p57 (KIP2), PP5, PLAC1, PLAC8 and PLAC9, CHL1, H119, HLA-C, Fra2, NDPK-A, the CAR protein, HASH2, human chorion gonadotropin alpha (hCGα), TFPI-2, annexin IV, cytokeratin-7, cytokeratin-8, cytokeratin-19, H19, IFG2, ESX1L, MASH2, ASH2, Stra13, FosB, Cyclin D1, GCM1, caspase-8, factor XIII, tapasin, Connexin 31, Connexin 43, HAND1, Syncytin, MMP9, APAF-1, caspase-3, caspase-9, FAS, Fas ligand, FLIP, AP-2γ, 313-HSD VI, CDX2, ERR2, PLX3, PTHrP, ASCL2, ID2, MET, TEF5, UPA, 11β-HSD2, c-ETs1, HMGI(Y), estrogen receptor, GemI, TTPA, 1503-7E, J42-4d, J2r(3), J2r(12), J2r(13), 305-4G, K1-1a, K2r / 1f(50), K2r / 1f(59), K(1)157-2A, K3r(HIGH)76, 597-10C, NT7-T3, N9r.Mf, 334-2C, O19r-T3, O1-1a, 332-9E, P60-1a, P1-1a, P3r(9), 305-9E, R5’-T3, R6r / 1-6H, 369-8G, U2f-T3, 305-6G, L15-1a, L21-1a, 252, 120r, clone-1, D19-2g, CD146, CD141, vimentin, VCAM, ICAM, VEGFR-1, VEGFR-2, VEGFR-3, ITGA5, ITGB5, CDH11, CDH3, CK1, CK2, CK3, CK4, CK5, CK6, CK7, CK8, CK9, CK10, CK13, CK14, CK15, CK16, CK17, CK18, CK19, CD9, CD39, CD200, EPHB4, EPCR, PAR-1, hPL, CSH2,WSGR Docket No. 62193-701.601 KISS1, GDF15, CRH, TFPI2, β-hCG, LOC90625, FN1, COL1A2, PSG9, HBE, AFP, GC, APOC3, SERPINC1, APOB, AHSG, HPX, CPB2, ITIH1, APOH, AMBP, Thy-1, CD90, thromobomodulin, Flt-1, EPCR, CDH11, CDH3, CD59, CD71, CD36, Glycophorin A, hexokinase 2, placental alkaline phosphatase, hyalouronic acid, BCL-2, VE-cadherin, PECAM- 1, α1-integrin, placental lactogen, PSG-1, α6 integrin, E-cadherin, MMP14, KCNQ4, CLDN6, F3, EPCAM, TROP2, CD147, fetal hemoglobin, embryonic hemoglobin, ITGA6, FSHR, CD45, LHCGR, CD270, Trop2, FOLR1, CD114, CSF3R, SLC40A1, CD156a, ERVW1, GPC3, TREML2, ALPP (866), CD163, CD144, ALPP (GM022), FOLR1, CD115, TSPAN1, SLC22A1, SCL22A11, EFNA1, SLC62A2, SLC6A2, NET, SLC40A1, CD71, CD166, TGF- beta2, human placental lactogen, c-erbB2, PAPPA2, PRG2, cytokeratin 7, beta-HCG, alpha- HCG, GCM1, syncytin, c-erbB2, leptin, INSL4, TGF-beta1, CSH1, KISS1, PEG10, PAGE4, p63, miR1246, miR1323, miR-512-3p, has-miR-516b-5p, miR-517-5p, has-miR525, miR-526b, ath-miR159a, RNU44, RNU48, U6, CDH9, HLA-G, MIH61, ALPP (8B6), CD49e, JAM2, transferrin receptor, glycophorin a (GPA), EGFR, thrombospondin receptor (CD36), CD34, HbF, HAE 9, FB3-2, H3-3, erythropoietin receptor, HBE, AFP, APOC3, SERPINC1, AMBP, CPB2, ITIH1, APOH, HPX, AHSG, APOB, J42-4-d, 2,3-biophosphoglycerate (BPG), carbonic anhydrase (CA), thymidine kinase (TK), MMP14, lectin, CD235a, carbohydrates, selectin, GPA, ANTIGEN-I, EpCAM, E-cadhrin, Muc-1, Hpl, CHS2, KISS1, GDF15, CRH, TFP12, CGB, LOC90625, FN-1, COL1A2, PSG9, PSG1, EGFR, CD105, cytokeratin, TLS1, CD147, CD141, PTPRN, BMPR2, TNFRSF19, STIM1, SELPLG, KLP-1, HECA452, GPR160, KLRD1, TNFSF18, LRP11, ABCA1, GPR176, SCARF, ESYT1, SCARB1, LTB4R, TLT2, CD141, CD90, Vimentin (Vim), VCAM, ICAM-1, VEGFR-1, Flt-1, VEGFR-1, VEGFR-2, VEGFR-3, PAI-1, EPCR, CD146, ITGA5, ITGB5, CDH11, CDH3, CD59, CSH-1, CSH-2, PSG2, PSG3, PSG11, HBG2, XAGE3, CSHL1, HBG1, ERVH48, phCG, any human orthologue of any of the foregoing, and any combination thereof.

[0118] In some embodiments, the subpopulation of fetal cells can have any combination of fetal biomarkers disclosed in the present disclosure.

[0119] In some embodiments, the different fetal cell types of the subpopulation of fetal cells can have expression of different fetal biomarkers.

[0120] In some embodiments, the fetal biomarkers of the present disclosure have human orthologues. Non-limiting examples of human orthologues of the fetal biomarkers include: AOC1, FAM236B, FAM236D, FAM236A, FAM236C, C11orf52, MAGEA10, A2M, A4GALT, AARD, ACVR2B, ADA, ADAMTS7, ADGRF5, ADM, ADRA2B, AFAP1L2, AIF1L, ALPL, APRT, ARHGAP22, ARHGAP42, BASP1, BCAR1, BEX1, BEX2, BGN,WSGR Docket No. 62193-701.601 BICC1, BMP1, BOK, BSG, KIAA1522, CALD1, CALR, CA4, CASKIN1, CAV1, CCL27, CD63, CDA, CDH3, CDH5, CDKN1C, CEACAM4, PSG3, PSG8, PSG1, PSG6, PSG7, PSG11, PSG2, PSG5, PSG4, PSG9, CHCHD10, CHRDL2, CITED1, CKB, CMTM5, COBL, COL4A1, COL4A2, COL5A2, COPZ2, COX6A1, CRIP2, CRYAA, CRYAB, CSRP1, CSTB, CYB5R3, CYP11A1, CYP17A1, CCN1, CYSTM1, DB1, DCN, DES, DLC1, DMKN, DNASE1L3, DSC2, DSP, DST, DSTN, DUSP9, EFS, EGFL7, EMP2, EPAS1, EPCAM, EPN3, EPS8L2, ESAM, EXOC3L4, FABP3, FABP6, FAM83B, FAM89A, FAM90A1, FAM90A26, FAM90A7, FAM90A22, FAM90A23, FAM90A14, FAM90A18, FAM90A16, FAM90A8, FAM90A17, FAM90A19, FAM90A9, FAM90A10, FBXL19, FDX1, FERMT2, FETUB, FKBP9, FMR1NB, FN1, FOXO4, FSCN1, FSTL3, GALE, GALK1, GHRH, GJB3, CT45A1, CT45A3, CT45A5, CT45A6, CT45A2, CT45A7, CT45A8, CT45A9, CT45A10, SAGE1, TMEM267, GPR50, H1- 0, HAND1, HDLBP, HMGN1, HOMER2, HRCT1, HSD17B2, HSPB1, HSPG2, HTATSF1, HTRA1, IGF2, IGF2BP1, IL33, ILDR2, INHBB, IRX3, KANK1, KLHL13, KRT18, KRT19, KRT7, KRT8, KRTDAP, LAMA1, LAMC2, LARS2, LDHB, LSOC1, LEPR, LGALS1, LGALS3, LMCD1, LMNA, LRRN4, LTBP4, MAGED1, MAGED2, MAP1B, MDK, MEIS2, MFSD2A, MGAT4B, MGST3, MMP1, MMP23B, MPDZ, MRGPRG, MT1G, MT2A, MUC1, MYOF, MYRF, NBL1, NENF, NFIB, NGEF, NID1, NOS2, NOSTRIN, NPC2, NPPC, NRK, NUP62CL, NXF5, NXF2, NXF2B, P4HB, ACP7, PARVA, PED10A, PDIA5, PDIA6, PEG10, PEG3, MPEG1, PHLDB2, PKP2, PLA2G5, PLAC1, PLET1, PLPP1, PODXL, PPIB, PPP2R2C, PRAMEF12, PRAMEF1, PRAMEF11, PRAMEF2, PRAMEF4, PRAMEF10, PRAMEF7, PRAMEF6, PRAMEF27, PRAMEF25, PRAMEF26, PRAMEF9, PRAMEF13, PRAMEF18, PRAMEF5, PRAMEF8, PRAMEF33, PRAMEF 15, PRAMEF14, PRAMEF19, PRAMEF17, PRAMEF20, PRDX4, PRKCD, PROCR, PROM1, PROM2, PRSS8, CEACAM4, PTOV1, NECTIN2, RAB15, RAB6B, RARRES2, RASD1, RBFOX2, RBP1, RGS17, RGS5, RHOC, RND2, RPN1, RPN2, S100A6, SBSN, SCT, SDC1, SEC61G, SEMA3E, SEMA3F, SERBINB6, SERBINB9, SERPINH1, SFN, SHROOM1, SIN3B, SLC30A2, SLC38A4, SLC6A2, SLCO2A1, SPARC, SSR4, STAR, STARD10, STRA6, SULT1E1, SYNGR1, TBRG1, TEX19, TFAP2C, TFF1, TFPI, TINAGL1, TMEM37, TMEM98, TNFRSF9, TPM1, TPM2, UACA, UQCRB, ATP5MK, VGF, FAM9A, FAM9B, FAM6C, C19orf33, ABHD6, ASNS, ATF5, ATP9A, BCAT1, CA2, CENPT, CGNL1, CTSL, CUL7, ELF3, FBXL19, GKAP1, GPX3, GPX8, H1FX, HDAC6, IFT43, JUP, LAMA5, LAMB2, OAF, PARM1, PAWR, PERP, PFPL, PHACTR1, PRNP, RGS16, RHOU, RRM2, SCIN, SERPINB6, SERPINB9, SERPINE1, SMIM1, SRC, TUBB3, and any combination thereof.WSGR Docket No. 62193-701.601

[0121] In some embodiments, the biomarkers for the subpopulation of fetal cells are Sct, H19, Krt8, Krt18, Tfpi, or Fthl17a. In some embodiments, the biomarker for the subpopulation of fetal cells is Sct. In some embodiments, the biomarker for the subpopulation of fetal cells is H19. In some embodiments, the biomarker for the subpopulation of fetal cells is Tfpi. In some embodiments, the biomarker for the subpopulation of fetal cells is Fthl17a.

[0122] In some embodiments, the subpopulation of fetal cells are stromal cells. Non-limiting examples of fetal biomarkers on fetal stromal cells include 4930486L24Rik, Cryab, Ctsl, Gpx3, Hsd3b6, Ly6c1, Mt1, Prl8a2, Sbsn, Htra1, Mfap5, Rrm2, Tfpi, Des, Dmkn, Mt2, Prnp, Cdh5, Col4a1, Cts7, H19, Hspb1, Lamb2, Lepr, Hspg2, Tfpi, Procr, Slc38a4, Mfap5, Des, Plet1, Cdh5, H19, Krt19, Plac1, Adgrf5, and Sct.

[0123] In some embodiments, the subpopulation of fetal cells are trophoblasts. Non-limiting examples of fetal biomarkers on fetal trophoblast cells include Rho9, Sct, H19, Prl2c5, Rhox6, Krt18, Krt8, Peg10, Tfpi, Ctsj, Hspb1, Cdkn1c, Prl2c3, Trap1a, Igf2, Fthl17a , Krt7, Krt19, Prl3b1, Prl7d1, Nrk, Rhox5, Sparc, Prl2c2, Procr, Prl7a1, Prl3d1, Ctsl, Ctsr, Crip2, Prl2b1, Sbsn, Cdh5, Hand1, Hsd17b2, Dusp9, Prl2a1, Rhox12, Ctsq, Serpinb9e, Fscn1, Plet1, Tnfrsf9, Cyp11a1, Nup62cl, 1600025M17Rik, Cited1, Prl4a1, Sdc1, Serpinb9b,Tinagl1, Bex1, Plac2, Car2, Gm7120, Cystm1, Ghrh, Gm9112, Nxf7, Rarres2, Serpinb9c, Slc38a4, Ceacam9, Fermt2, Serpinb9d, Syngr1, 3830417A13Rik, Mfap5, Serpinb9g, Klhl13, Map1b, Psg22, Rbp1, Ldoc1, Prl8a2, Serpinb9f, Adgrf5, Fnd3c2, Parva, Prl2c1, Cyp17a1, Lepr, Prl3d2, Prl8a9, Psg29, Cts6, Gpx3, Mt1, Prl3d3, Prl8a1, Col4a1, Fmr1nb, Homer2, Perp, Cts7, Prl5a1, Serpine1, Slc30a2, Tfap2c, Hsd3b6, Igf2bp1, Oaf, Pramef12, Shroom1, Tpbpa, 1700011N02Rik, Sult1e1, 4930486L24Rik, Bcat1, Elf2, Ly6C1, Mt2, Nppc, Pfpl, Rrm2, Hspg2, Mmp1a, Phactr1, Prl8a2, Vgf, Cdh3, Cryab, Prnp, Zfp36l3, 1600015l10Rik, Prl2c5, Rhox9, Krt18, Sct, Serpinb9d, Krt14, Hand1, Serpinb9g, Slc38a4, Cyp17a1, Nppc, Hsd3b6, Prl7a1, Prl3d1, Rbp1, Bcat1, Prl3b1, Ctsj, Fthl17a, Hsd17b2, Nppc, Cdkn1c, Syngr1, Ctsj, Cdh5, Tfpi, Sdc1, Hspg2, Lepr, Plac1, Cryab, Procr, Plet1, Adgrf5, Hspg2, Hspb1, Lepr, Tnfrsf9, Sdc1, Nup62cl, Ctsr, Zfp36l3, Map1b, Igf2, Trap1a, Nrk, and Chrdl2.

[0124] In some embodiments, the subpopulation of fetal cells are early gestation trophoblasts (e.g., early gestation trophoblasts). Non-limiting examples fetal biomarkers on early gestation trophoblasts include Prl3d1, H19, Krt18, Prl7a1, Sct, Krt8, Prl2c3, Rhox9, Rhox6, Prl2c5, Cdkn1c, Krt19, Prl2c2, Hspb1, Krt7, Igf2, Prl4a1, Sbsn, Prl2a1, Cyp11a1, Hand1, Peg10, Procr, Trap1a, Ctsl, Tfpi, Cdh5, Psg22, Sdc1, Serpinb9b, Plac1, Rhox5, Dusp9, Gm7120, Nrk, Serpinb9e, Plet1, Sparc, Cystm1, Fscn1, Fthl17a, Prl3d2, Prl3d3, Prl8a1, Crip2, Ctsj, Parva, Psg29, Serpinb9c, Serpinb9d, Tnfrsf9, Hsd17b2, Nup62cl, Tinagl1, 1600025M17Rik,WSGR Docket No. 62193-701.601 Serpinb9g, 3830417A13Rik, Car2, Nxf7, Prl7d1, Prl8a9, Rhox12, Serpinb9f, Cyp17a1, Homer2, Ldoc1, Prl2c1, Prl5a1, Adgrf5, Hsd3b6, Igf2bp1, Rbp1, Col4a1, Fermt2, Map1b, Prl2b1, Prl3b1, Tfap2c, Serpine1, Bcat1, Cryab, Fmr1nb, Nppc, Oaf, Sult1e1, Syngr1, Vgf, Hspg2, Mt2, Perp, Rrm2, Slc30a2, 1600015l10Rik, Ceacam9, Chrdl2, Cited1, Ctsr, Gm9112, Mt1, Pramef12, Slc38a4, Dst, Map1b, Gm7120, Map1b, Rere, Prl3d1, Prl5a1, Krt7, Prl8a2, Prl7a1, Rbp1, Serpine1, Procr, Serpinb9b, Sdc1, Prnp, Fscn1, Psg22, Fthl17a, Krt14, Cdh3, Fnd3c2, 1700011M02Rik, Prl3d2, Gm9112, Ldoc1, 3830417A13Rik, Serpinb9f, Igf2bp1, Sult1e1, Pramef12, Zfp3613, Rhox5, Rhox12, Plac1, Serpinb9g, Prl2cl, Adgrf5, Slc38a4, Rrm2, Cryab, Htra1, Des, Mfap5, Dmkn, Ces1a, Plac1, Tfpi,Hspg2, Plet1, Cdh5, Lepr, Procr, Sdc1, Chrdl2, Prl5a1, Krt14, Cdh3, Fnd3c2, Prl3d2, Ldoc1, Pramef12, Psg29, Prl3d3, Psg22, Prl8a1, Parva, Slc30a2, Ghrh, Mmp1a, Hsd3b6, Prl5a1, Prl3d2, Trap1a, Nrk, Plet1, Fthl17a, Plac1, Tfpi, Hspg2, Cdh5, Lepr, Procr, Cryab, Sdc1, Hsd3b6, Des, Mfap5, Htra1, Dmkn, and Tpbpa.

[0125] In some embodiments, the subpopulation of fetal cells comprises later gestation trophoblasts (e.g., late gestation trophoblasts). Non-limiting examples of fetal biomarkers on later gestation trophoblasts include Ctsq, Ctsr, Prl3b1, Prl7d1, Prl3d1, Cyp11a1,Cdh5, Hand1, Cdh3, 1600025M17Rik, Rhox5, Fermt2, Serpinb9d, Serpinb9f, 3830417A13Rik, Fnd3c2, Pramef12, Serpinb9g, Nup62cl, Prl2cl, Fnd3c2, 170011M02Rik, 3830417A13Rik, Nup62cl, Adgrf5, Plac1, Tpbpa, Pel3d3, Tfap2c, Serpinb9e, Psg29, Parva, Serpinb9c, Prl8a9, Psg22, Slc30a2, Hsd3b6, Prl8a1, Nxf7, Prl2a1, Htra1, Prl4a1, Dusp9, Tinagl1, Plet1, Pel7a1, Prl7d1, Ctsj, Sbsn,Prpf4b, Erdr1, Gm7120, Prl3d2, Igf2bp1, Ldoc1, Sult1e1, Krt14, Feng3c2, Fthl17a, Rhox12, Adgrf5, Fermt2, Prpf4b, Ctsl, Prl2b1, Zfp3613, Fnd3c2, Rhox6, Rhox9, Sct, Ctsj, Prl2c5, H19, Prl2b1, Sparc, Trap1a, Peg10, Rhox6, Tfpi, Bex1, Krt18, Cited1, Cts6, Ghrh, Hsd17b2, Hspb1, Rarres2, Crip2, Krt8, Lepr, Mfap5, Mt1, Nrk, Prl3d2, Procr, Fthl17a, Prl2c3, Slc38a4, Fermt2, Gm9112, Krt19, Cdkn1c, Krt7, Map1b, Rhox12, Igf2, Sbsn, and Syngr1.

[0126] In some embodiments, the fetal cell biomarker is an extracellular biomarker. In some embodiments, the fetal cell biomarker is an extracellular protein. In some embodiments, the extracellular protein is a cell surface protein. In some embodiments, the cell surface protein is a cell-surface receptor. In some embodiments, the extracellular protein is a soluble cell surface protein. In some embodiments, the extracellular protein is a transmembrane protein. In some embodiments, the extracellular protein is a transmembrane channel protein.

[0127] In some embodiments, the fetal cell biomarker is an intracellular biomarker. In some embodiments, the fetal cell biomarker is an intracellular protein. In some embodiments, the intracellular protein is an enzyme. In some embodiments, the intracellular protein is a phosphoprotein. In some embodiments, the intracellular protein is a growth factor. In someWSGR Docket No. 62193-701.601 embodiments, the intracellular protein is a structural protein. In some embodiments, the fetal cell biomarker is a nucleic acid molecule. In some embodiments, the nucleic acid molecule is DNA. In some embodiments, the nucleic acid molecule is RNA. In some embodiments, the RNA is mRNA. In some embodiments, the RNA is rRNA. In some embodiments, the RNA is lncRNA. In some embodiments, the RNA is miRNA. In some embodiments, the RNA is siRNA.

[0128] In some embodiments, the subpopulation of maternal cells has one or more maternal cell biomarkers. In some embodiments, the maternal cell biomarker is a cell-surface protein. In some embodiments, the maternal cell biomarkers is a cell-surface receptor. In some embodiments, the maternal cell biomarkers is a nucleic acid biomarker. In some embodiments, the nucleic acid biomarker is a RNA biomarker. In some embodiments, the nucleic acid biomarker is a DNA biomarker. In some embodiments, the biomarkers for the subpopulation of maternal cells are different from one or more fetal cell biomarkers. Non-limiting examples of maternal cell biomarkers include Coro1a, Rac2, Malat1, Laptm5, H2-D1, B2m, Cytip, Alox5ap, Cd52, Junb, Cyba, Fyb, Btg1, Srgn, Actb, Klf2, Mcl1, H3f3a, Hmha1, Tmsb4x, Cd53, Ptprc, Fxyd5, Myl12b, Tyrobp, Gmfg, Fcer1g, Ier2, H2-K1, Itm2b, Jund, Rasgrp2, Nfkbia, Itgb2, Pfn1, Ifngr1, Ltb, Pim1, Ssh2, Rbm39, Sell, Trbc2, Msn, Ddx5, Stk17b, Lyz2, Cd37, Csf3r, Hcst, Ifitm2, Pnrc1, Rap1b, Meat1, Fth1, Cd3d, Arhgdib, Pglyrp1, H3f3b, Ms4a6b, Akap13, S100a9, Lyn, Sorl1, Mbnl1, Cd44, Ier5, Lsp1, Cdc42, Aes, Ms4a6c, Msrb1, Gdpd3, Btg2, Sat1, Arhgap30, Lrg1, Itga4, Oaz1, Rhoa, Eif1, Zfp36l2, Selplg, Cd3g, Zfp36, Arhgap15, Syk, Arpc2, Sf3b1, Ighm, Psmb9, Gnai2, Actr3, Lst1, Il1b, Satb1, Amp32a, Foxp1, Lcp1, Wfdc17, Taldo1, Slfn2, Rgs10, Ccnd3, Ncor1, Cd3e, Vim, Cebpb, Tpm3, Adgre5, S100a8, Ccl6, Ptpn18, Napsa, Rps9, C5ar1, Hnrnpf, Iqgap1, Ly6e, Klf13, Ifitm3, Psmb8, H2-Q4, Gsr, Hmgb2, Fos, Sh3bgrl3, Dusp1, Spi1, Cd84, Tspo, Lef1, Cyp4f18, Atp11b, Ccr1, Gpx1, Cd47, Samhd1, AB124611, Trac, AI467606, Shisa5, Myl6, Tspan13, Cfl1, Gm5150, Itgal, Calm2, Plek, Ucp2, Tsc22d3, Ikzf1, Grk6, Gimap1, Ybx1, Gm8797, Clic1, Rara, Limd2, Ifi27l2a, Ubb, Gngt2, Cd14, Pla2g7, Ets1, Tgfb1, Actg1, Il17ra, Celf2, Srsf5, Gimap6, Tmem50a, Ctss, Ube2d3, Tln1, Apbb1ip, Gabarap, Arpc5, Kmt2e, Klf3, Cap1, Itgam, Emp3, Clk1, Ogt, Crlf3, Samsn1, Stk38, Tax1bp1, Lat, Clec4e, Cdkn2d, Rel, Srrm2, Smc6, Morf4l1, Rsrp1, Ptpn1, Rgs2, Fmnl1, Gpsm3, Klf6, Fcgr3, Mef2c, Birc2, Kdm6b, Tra2b, Ptprcap, Son, Rassf5, Rpl22, Gimap3, Hnrnpa2b1, Lbr, Lck, Calm1, Cdkn1b, Ccr7, Ypel3, Ptpn6, Cd74, Ppp2r5a, Cd9, Rcsd1, Svil, Hmgb1, Rhog, Hacd4, 9930111J21Rik2, Stat1, Ncf4, Fam105a, Rsbn1l, Hp, Ankrd11, Emb, Dusp2, Pfdn5, Txnip, Psme1, Slfn1, Ostf1, Jak1, Sirpa, Rap1a, Fam49b, Gda, Picalm, Ywhaz, Pde4b, Dazap2, Tsc22d4, Pkm, Slc25a5, Kdm7a, Lcp2, Fus, H2-Q7, Mxd1, Glipr1, Bnip3l, Luc7l2, Tnfaip8l2, Pdcd4, Map1lc3b, Plbd1, Skap1, Unc93b1, Ncf2, Rnf149, Bin2, Mycbp2, Cnp, S100a10, Ubc,WSGR Docket No. 62193-701.601 Cd300ld, Fam32a, Ptp4a2, Adrb2, Prdx5, Crip1, Sna32, Csk, Tnrc6b, Arhgef1, Pou2f2, Hspa8, Pilra, Bcl11b, Zfp292, Cnn2, Aldh2, Ywhah, Zbtb7a, Pycard, Cotl1, Retnlg, Sp100, Irf2, Fgr, Igsf6, Ablim1, Fam65b, Fermt3, Tmbim6, Tpd52, Scp2, Ube2b, Notch2, Gnb2, Il7r, Capzb, Jun, Ptma, 2810474O19Rik, Prr13, Mrpl33, Ppp1r15a, Lilrb4a, Zyx, St8sia4, Arpc3, H2afz, S100a11, Pag1, Irf1, Tm6sf1, Rsf1, Frina, Rasa3, Smap2, Dok3, Skap2, Add3, Stx11, Mkrn1, Hck, Ccnl1, Ppp1ca, R3hdm4, Cd48, Chd2, Rgs19, Cth4, Atp2b1, Peli1, Fam107b, Ptbp3, Adipor1, Serp1, Pak2, Hspa5, Tpr, Zc3hav1, Sdcbp, Dock2, Rnf130, Ftl1, B4galnt1, Cd300a, Hcls1, Anxa1, Cst3, Ankrd44, Elf1, Prpf38b, Atp6v1g1, Snx20, Il6ra, Whsc1l1, Tmem71, Stk4, Sla, Mbp, Dnaja1, Vgll4, Capza2, Ldha, Pyhin1, Rapgef6, Hdc, Fli1, Rbm25, H2afy, Prdx6, Rac1, Stap1, Supt4a, Hnrnpk, Ubl5, Cdk2ap2, Ppp1r2, Psenen, Bcl2, Tprgl, Arpc1b, Cd69, Phf20l1, Ppt1, Rhoh, Coro1a, Cytip, Malat1, Rac2, Cd52, Laptm5, B2m, H2-D1, Cyba, Ptprc, Btg1, Gmfg, Junb, Srgn, Fxyd5, Itgb2, Ltb, Tyrobp, Mcl1, Cd53, Klf2, Fcer1g, H3f3a, Hmha1, Itga4, Alox5ap, Ptpn18, Myl12b, Cd3d, Spi1, Ier2, Ccl6, Ms4a6b, Msn, Selplg, Pglyrp1, S100a9, Lyn, Slfn2, H2-K1, Ier5, Cyp4f18, Csf3r, Hp, Lst1, Jund, Fyb, Rap1b, Pim1, Il1b, Lyz2, Stk17b, Cd3e, Gpsm3, Lef1, Sell, Hcst, Gm5150, Ms4a6c, Fmnl1, Adgre5, Arhgap30, Lrg1, 9930111J21Rik2, Unc93b1, Rbm39, Pnrc1, Ptpn6, Trac, Akap13, Actr3, Ifitm2, Crlf3, Msrb1, Samsn1, Ets1, Lck, Pla2g7, Zfp36l2, Lat, Ncor1, Sorl1, Ccnd3, Aes, Rasgrp2, Samhd1, Psmb8, S100a8, Wfdc17, Ctss, Neat1, Arhgap15, Ncf4, Lcp1, AB124611, Ssh2, Btg2, March1, Snx20, Fam105a, Zfp36, C5ar1, Tspan13, Hck, Apbb1ip, Mbnl1, Cd37, Limd2, Skap1, Iqgap1, Cybb, Lcp2, Ly6e, Slfn1, Bcl11b, Syk, Sf3b1, Ccr1, Ddx5, Stk38, Sat1, Anp32a, Dock2, Cd74, Satb1, Ighm, Foxp1, Tspo, Klf13, Ptpn1, AI467606, Nfam1, Cap1, Hnrnpf, Shisa5, Fgr, Rhog, Ifngr1, Psmb9, Rhoh, Lilrb4a, Plek, Rasa3, Sla, Atp11b, Fam65b, Gimap1, Hmgb2, Ypel3, Tnfaip8l2, Pirb, Rgs2, Tmem50a, Fcgr3, Cd84, Kmt2e, Mpeg1, Retnlg, Ikzf1, Il6ra, Prdx5, Ostf1, Cd14, Itgam, Cd300ld, Rnf149, Cd79b, Dusp1, Tsc22d4, Ifi27l2a, Il17ra, Sirpb1c, Klf3, Celf2, Clic1, Cebpb, Grk6, Pira2, Grap, Ppp2r5a, Fam49b, Cd47, Tgfb1, Wfdc21, Hcar2, Ncf2, Cd300a, Ms4a4b, Itgal, Trbc2, Dusp2, Tra2b, Pilra, Cyth4, Hcls1, Pyhin1, Son, Tgfbi, Tspan32, Vav1, Cd3g, Irf1, Dok3, Cxcr2, Ifitm3, Rara, 2810474O19Rik, Cd33, Adrb2, Snap23, Fos, B4galnt1, Lilr4b, Tpm3, Prkcq, Cd44, Rcsd1, Sp100, S1pr1, Cdk2ap2, Cd247, Cotl1, Lrrfip1, Slc11a1, Srsf5, Cnp, Klf6, Clec4e, Inpp5d, Glipr1, Lpcat2, Fermt3, Clec4a3, Fus, Itgb7, Tbc1d10c, Napsa, Pld4, Add3, Jak1, Aldh2, Cdkn1b, Gsr, Myo1f, Ccr7, Mxd1, St8sia4, Gimap3, Arpc5, Ly6d, Rassf5, Ifitm1, Arhgef1, Gdpd3, Ifitm6, Smap2, Fam107b, Cd224, Mcemp1, Bin2, Hdc, Fam32a, Pou2f2, Kdm6b, Cdkn2d, Vim F630028O10Rik, H2-Q4, Pag1, Rnf130, Smpdl3a, Tpd52, Tmcc1, Grina, Fgl2, Csk, Trbc1, Cd27, D1Ertd622e, Rasal3, Gabarap, Cd28, Ipcef1, Rgs19, Bcl2, 4930523C07Rik, Slc25a5, Rapgef6, Ppt1, Gngt2, Irf2,WSGR Docket No. 62193-701.601 Psme1, Clec4d, Slc15a3, Pilrb2, Andrk11, Peli1, Tsc22d3, Bank1, Iglc2, Il7r, Stfa2l1, Stat1, Igsf6, Tax1bp1, Nedd9, Tm6sf1, Ly86, Tprgl, 2310001H17Rik, H2-DMa, H2-DMb2, Sirpb1b, Srrm2, Stk4, H2-T23, Abcg1, Klhl6, Ablim1, Evi2a, Bnip3l, Arhgap25, Lrrc25, Ogf rl1, Emp3, Clk1, Mkrn1, Ggnt2, Hacd4, Picalm, Adipor1, Man2b1, Cd48, Lyl1, Sp140, Sephs2, Lbr, Cd69, Phf20l1, Scp2, Ms4a1, Cdc42se1, Rgs10, Sirpa, Stap1, Ddx6, Ppp1r2, Abi3, Acap1, Csf2rb, Tnfrsf18, Alox12, Ebf1, Luc7l2, Flna, Ikzf3, Sept1, Arid4a, Pi16, Gimap6, Dazap2, Ppp1r16b, Lyst, Adrbk1, Nlrp3, Ptprcap, Pycard, Themis2, Lbh, Mycbp2, Rel, Apobec1, Cd2, Chd2, AF251705, Trem1, Dennd1c, Ram189b, Ncf1, Tnfaip2, Pde4b, Ccnl1, Fcgr4, Skap2, Elf1, Il2rg, Slc7a11, Trim30b, Smchd1, H2-Q7, Rasgrp1, Ptpn22, Mrgpra2b, Ptpn22, Mrgpra2b, Cmip, Dgka, 1600010M07Rik, Igkc, Slc2a6, Atg3, Arl4c, Il16, Btk, Il21r, Fli1, Gda, Mbd2, Fam134b, Myo1g, Pygl, Ptpre, Kdm7a, Ctse, Plbd1, Map3k3, Akna, Mmp8, Ywhaz, H2afy, Svil, Mfsd6, Emilin2, Hnrnpa0, Psenen, Pdcd4, Cd5, Chil1, Pnpla2, Macf1, Gm8369, Nfatc1, Plcg2, Rhof, Gm9733, Map1lc3b, Capzb, Ube2b, Ccpg1, Il1f9, Xist, Gimap5, Rsbn1l, Cbl, Cd300lb, Siglece, Slco3a1, Supt4a, Mbp, Ms4a4c, Cd79a, Nrros, Gimap7, Birc2, Ccdc12, Mapk14, Tcf7, Whsc1l1, I830077J02Rik, Rbm25, Xrn2, Jakmip1, Dock10, Vsir, Siglecg, Ramp1, Cd6, Phip, Ifi203, Tpr, Kpna4, Ptafr, Sh3bp5, Itk, Tnrc6c, Notch2, Myd88, Nipbl, Prkcd, Slc2a3, Bcl10, Il10ra, Dhrs7, Clec4a1, S1pr4, Cd300lf, Ogt, Lrmp, Ttc7, Gm43603, Treml1, Bcl2a1b, Marcks, Mier1, Vasp, Clec4a2, Gpcpd1, Traf3ip3, Prpf38b, Runx3, R3hdm4, Lfng, 1110008F13Rik, Slamf6, Pdcd10, Stx11, Srsf11, Gp5, Hnrnpl, BC028528, Gm26740, Pid1, Nfe2, Mzb1, Lilra6, Snca, Parp14, Vgll4, Socs3, Sh2d1a, Dapl1, Gpr183, AW011738, Hbb-bs, Lmnb1, Strbp, H2- Aa, Il18rap, Hipk1, Stag2, Orai1, Smc6, Stk10, Fam111, Treml4, Ctage5, Ebi3, Ap1s2, Neurl3, Trib2, Zc3hav1, Baz1a, Hectd1, Sec11c, Tnrc6b, Amica1, Prkd2, Rnase6, Evl, Vav3, Rab27a, Trim30a, Adam8, Phf3, Gpr132, Sirt7, Tcp11l2, Zcchc7, Itpr2, Bptf, Ace, Icam2, Slfn4, Fcmr, Prkar1a, Asap1, Clec2i, Prr13, Mef2c, Kansl1, Slc40a1, Vps37b, Sdcbp, Cd8b1, Phf11b, Zeb2, Sfpq, Ccl3, Anxa1, Pik3cd, Serp1, Arrb2, Ccdc88b, Ctsc, Atf7ip, Treml2, Nfkb1, Vamp8, Slc16a3, Kif2a, Lmo2, Capza2, Tbc1d1, Gimap8, Rassf3, Ifngr2, Ltb4r1, Trem3, Pf4, Clec2d, Pten, Cd226, H2-Ab1, Sgk1, Zcchc11, Lcn2, Rgs14, Ptp4a2, Tmem156, Mctp1, H2-Q6, N4bp1, Arhgap9, Birc3, Txk, Nptn, Psme2, Arl6ip5, Pstpip1, Tuba4a, Sertad1, Diaph1, Tnfrsf13b, Batf, AW112010, Il10rb, Arl5c, Cxcl2, Ccdc88c, Cdk9, Sh3bp1, Gimap4, Rnf144a, Bcl11a, Rassf2, Gramd1a, Nadk, Tnfrsf13c, Tra2a, Ggnbp2, Isg15, Def6, Stat4, Prrc2c, Gm15987, Card19, Clec12a, Gsap, Mfng, BC021614, Cmtm6, Cpr65, Tctex1d2, Cntrl, Tacc1, Zfp292, Sh3kbp1, Ifi47, Gpr18, Syf2, Twistnb, Rinl, Gp9, Aim2, Ctcf, Cd300e, Dck, B430306N03Rik, Otulin, Pip4k2a, Max, Tnfsf14, Krit1, Tmpo, Tnf, Ubl3, Csnk1a1, Fpr2, Cecam1, Bmx, Vrk1, Prkcb, Pfkfb4, G3bp2, Abtb1, Epsti1, Rin3, Itpkb, Gimap9, Adgre4, Ms4a6d, Ly9, Tap2, Prkar2b,WSGR Docket No. 62193-701.601 Sf3b6, Fam78a, Adhb8, 1700097N02Rik, Zdhhc18, Atp1a3, Ppp1r10, Ifi30, Mcmbp, Twf2, C130050O18Rik, Kras, Arhgap4, Tmem71, Alcam, Notch1, Morc3, Sun2, Haus8, Rsrc2, Cdc42ep3, Ikbkb, Chd7, Arl11, Clec4n, Cxcr4, Stk24, Nfkbiz, Serinc3, Timm10b, Ctsw, Rchy1, Tap1, Maf1, Rbm5, Sema4a, Mat2b, Cth1, Nfkbid, Cd4, Ly6c2, Mef 2a, March2, Grap2, Avl9, Fcer2a, Cd24a, Hba-a1, Bcl2a1a, Arf6, Rnf13, Blnk, Il27ra, Kcnn4, Hn1, Fpr1, Sash3, Atp8b4, Sfxn5, Gpr141, Rsu1, Prr5l, Scnn1a, Tmem243, Fcrl1, Pik3r1, Nlrp12, Milr1, THBD, F3, FLT3, MCAM, CD141, CD135, CD142 / F3, CD146, CD33, CD68, CD144, CDH5, CD62E, CD45, CD90, SELE, GAP / GLA5, ITGA5, KCNK3 / TASK-1, CNTFR, JAM-1 (f11r), SDC4 / Syndecan 4, ACVR2B, ACVR2A, CD44v6+CD44v4 / 5, STX1A, TEK / Tie2, CD171, L1CAM, NCAM1 / CD56, ALPP, ALPPL2, MMP23A / MMP23B, CD14, CD4, CD8, CD3, CD19, CD32, CD16, HLA-A, HLA-B, CD15 FSHR, CD45, LHCGR, CD270, Trop2, FOLR1, CD114, CSF3R, SLC40A1, CD156a, ERVW1, GPC3, TREML2, ALPP (866), CD163, CD144, ALPP (GM022), FOLR1, CD115, TSPAN1, SLC22A1, EFNA1, SLC62A2, SLC40A1, CD71, CD166, CDH12, HLA-A, HLA-B, PRSS8, CD45, CD230, ALPP (8B6), CD49e, JAM2, TREML2, MMP14, lectin, CD47, CD45, CD35, CD12, CD14, CD32, CD235a, carbohydrates, selectin, GPA, ANTIGEN-I, EpCAM, E-cadherin, Muc-1, Hpl, CHS2, KISS1, GDF15, CRH, TFP12, CGB, LOC90625, FN-1, COL1A2, PSG9, PSG1, CD105, TLS1, CD147, CD15, TNFRSF19, STIM1, CD19, CD14, any human orthologue of any of the foregoing, and any combination thereof.

[0129] In some embodiments, the maternal biomarkers of the present disclosure have human orthologues. Non-limiting examples of human orthologues of the maternal biomarkers include: CYBA, CORO1A, CYTIP, LAPTM5, CD53, RAC2, CD52, PTPRC, FCER1G, ARHGAP45, GPM6A, MAF, C3, CFH, EFEMP1, BICC1, COL1A2, COL1A1, B2M, BTG1, GMFG, JUNB, SRGN, ITGB2, LTB, TYROBP, MCL1, KLF2, NFKBIA, H3F3A, ITGA4, ALOX5AP, PTPN18, CD3D, SPI1, IER2, CCL15-CCL14, CCL14, CCL15, CCL23, MS4A6A, MS4A6E, MSN, SELPLG, ACTB, PGLYRP1, TMSB4Y, LYN, SLFN12, SLFN12L, IER5, CYP4F3, CSF3R, HP, HPR, LST1, ITM2B, JUND, FYB1, LSP1, RAP1B, PFN1, PIM1, IL1B, STK17B, CD3E, GPSM3, LEF1, SELL, HCST, AL049634.2, SIRPB1, SIRPG, SIRPA, MS4A6A, MS4A6E, FMNL1, ADGRE5, ARHGAP30, LRG1, UNC93B1, RBM39, PNRC1, PTPN6, AKAP13, ACTR3, IFITM2, IFITM1, IFITM3, ARHGDIB, CRLF3, MSRB1, SAMSN1, ETS1, LCK, PLA2G7, ZFP36L2, LAT, NCOR1, SORL1, CCND3, AES, RASGRP2, SAMHD1, PSMB8, S100A8, CTSS, ARHGAP15, NCF4, LCP1, C19orf38, SSH2, BTG2, MARCH1, GNAI2, SNX20, FAM105A, ZFP36, C5AR1, TSPAN13, HCK, APBB1IP, MBNL1, CD37, LIMD2, SKAP1, IQGAP1, CYBB, LCP2, EMB, LY6E, SLFN12, SLFN12L, BCL11B, SYK,WSGR Docket No. 62193-701.601 SF3B1, CCR1, DDX5, STK38, SAT1, ANP32D, ANP32A, TALDO1, CDC42, DOCK2, CD74, SATB1, IGHM, TSPO, KLF13, H3F3B, PTPN1, C16orf54, NFAM1, CAP1, HNRNPF, SHISA5, FGR, RHOG, IFNGR1, PSMB9, RHOH, PLEK, RASA3, SLA, ATP11B, RIPOR2, SH3BGRL3, GIMAP1, HMGB2, RHOA, YPEL3, TNFAIP8L2, LILRB3, LILRA6, LILRB5, LILRA4, ARPC2, RGS2, TMEM50A, FCGR2A, FCGR2C, FCGR2B, CD84, KMT2E, MPEG1, RETNLB, TXNIP, IKZF1, IL6R, PRDX5, OSTF1, CD14, ITGAM, CD300LF, RNF149, CD79B, DUSP1, TSC22D4, IL17RA, AL049634.2, KLF3, CELF2, CLIC1, CEBPB, GPX1, GRK6, LILRA5, GRAP, GRAPL, PPP2R5A, FAM49B, CD47, TGFB1, HCAR2, HCAR3, NCF2, CD300A, CD300LD, ITGAL, TRBC1, TRBC2, OAZ1, DUSP2, TRA2B, CFL1, CYTH4, ANKRD44, HCLS1, SON, TGFBI, TSPAN32, VAV1, CD3G, IRF1, MRPL33, DOK3, CXCR2, IFITM1, IFITM3, IFITM2, RARA, KIAA1551, CD33, SIGLEC6, ADRB2, SNAP23, FOS, B4GALNT1, PRKCQ, CD44, RCSD1, UCP2, SP140, S1PR1, TLN1, CDK2AP2, CD247, COTL1, LRRFIP1, SLC11A1, SRSF5, CNP, CNN2, KLF6, CLEC4E, INPP5D, GLIPR1, LPCAT2, FERMT3, CLEC4C, FUS, ITGB7, TBC1D10C, NAPSA, PLD4, ADD3, JAK1, AC002996.1, ALDH2, CDKN1B, GSR, MYO1F, CCR7, MXD1, ST8SIA4, GIMAP1-GIMAP5, GIMAP1, GIMAP5, ARPC5, LY6D, RASSF5, IFITM1, IFITM2, IFITM3, ARHGEF1, GDPD3, SMAP2, FAM107B, MCEMP1, BIN2, HDC, FAM32A, POU2F2, KDM6B, CDKN2D, VIM, PAG1, RNF130, SMPDL3A, AC036214.3, TPD52, TMCC1, GRINA, FGL2, CSK, TRBC1, TRBC2, CD27, MACIR, RASAL3, GABARAP, CD28, IPCEF1, RGS19, BCL2, KIAA0040, SLC25A5, RAPGEF6, AC008695.1, PPT1, GNGT2, IRF2, PSME1, CLEC4D, SLC15A3, ANKRD11, PELI1, TSC22D3, RSRP1, YBX1, BANK1, IGLL1, IL7R, CTSA, STAT1, IGSF6, TAX1BP1, NEDD9, TM6SF1, LY86, TRPG1L, HLA- DMA, HLA-DMB, SRRM2, STK4, ABCG1, KLHL6, ABLIM1, RAP1A, EVI2A, BNIP3L, ARHGAP35, LRRC25, OGFRL1, EMP3, CLK1, MKRN1, GCNT2, HACD4, PICALM, UBE2D3, ADIPOR1, MAN2B1, CD48, LYL1, PAK2, SEPHS2, LBR, CD69, PHF20L1, SCP2, MS4A1, CDC42SE1, RGS10, STAP1, DDX6, PPP1R2, PPP1R2B, ABI3, ACAP1, PKM, CSF2RB, TNFRSF18, ALOX12, EBF1, LUC7L2, FLNA, IKZF3, PRDX6, SEPTIN1, ARID4A, PI16, GIMAP6, DAZAP2, PPP1R16B, LYST, GRK2, NLRP3, HNRNPA2B1, PTPRCAP, PYCARD, THEMIS2, LBH, MYCBP2, REL, APOBEC1, CD2, CHD2, TREM1, DENND1C, ENTREP3, NCF1, TNFAIP2, PDE4B, ATP6V1G1, CCNL1, FCGR3A, FCGR3B, SKAP2, ELF1, IL2RG, SLC7A11, SMCHD1, CD9, RASGRP1, PBXIP1, PTPN22, MRGPRX3, MRGPRX4, MRGPRX1, MRGPRX2, CMIP, DGKA, IGKC, SLC2A6, ATG3, ARL4C, IL16, BTK, IL21R, PPP1CA, FLI1, GDA, MBD2, MYO1G, PYGL, SCAND1, PTPRE, KDM7A, CTSE, PLBD1, MAP3K3, AKNA, MMP8, YWHAZ, MACROH2A1, SVIL,WSGR Docket No. 62193-701.601 H2AJ, MFSD6, ZYX, EMILIN2, HNRNPA0, PSENEN, PDCD4, CD5, CHI3L1, PNPLA2, MACF1, NFATC1, PLCG2, RHOF, CALM2, ROCK1, MAP1LC3B2, MAP1LC3B, CAPZB, UBE2B, CCPG1, IL36G, EIF1, GIMAP1-GIMAP5, GIMAP1, GIMAP5, MORF4L1, RSBN1L, CBL, CD300LB, SIGLEC9, SIGLEC7, SIGLEC8, SIGLEC12, SLCO3A1, SUPT4H1, MBP, PPP1R15A, CD79A, NRROS, PPP1R18, PTBP3, GIMAP7, BIRC2, CCDC12, MAPK14, TCF7, C1orf162, RBM25, XRN2, JAKMIP1, DOCK10, VSIR, SIGLEC11, SIGLEC10, RAMP1, CD6, PHIP, TPR, KPNA4, PTAFR, SH3BP5, ITK, TNRC6C, NOTCH2, MYD88, NIPBL, PRKCD, BCL10, IL10RA, DHRS7, CLEC4A, S1PR4, CD300LF, OGT, IRAG2, TTC7A, TREML1, FAU, BCL2A1, MARCKS, MIER1, VASP, CLEC4C, GPCPD1, TRAF3IP3, RUNX3, R3HDM4, LFNG, SLAMF6, PDCD10, STX11, SRSF11, GP5, HNRNPL, C1orf54, PID1, NFE2, MZB1, LILRA5, SCNA, PFDN5, PARP14, VGLL4, ENO1, ARL6IP1, SOCS3, SH2D1A, DAPL1, GPR183, LMNB1, STRBP, IL18RAP, HIPK1, STAG2, ORAI1, SMC6, STK10, FAM111A, GNB2, TREML4, EBI3, AP1S2, ARPC1B, NEURL3, TRIB2, ZC3HAV1, BAZ1A, HECTD1, SEC11C, TNRC6B, PRKD2, RNASE6, EVL, VAV3, RAB27A, TRIM5, ADAM8, PHF3, SIRT7, TCP11L2, ZCCHC7, ATP2B1, ITPR2, BPTF, ACE, ICAM2, SLFN12, SLFN12L, FCMR, PRKAR1A, ASAP1, CLEC2D, MEF2C, KANSL1, SLC40A1, VPS37B, SDCBP, CD8B, CD8B2, PHF11, ZEB2, SFPQ, CCL18, CCL3, CCL3L1, ANXA1, YWHAH, HMGB1, PIK3CD, SERP1, ARRB2, CCDC88B, CTSC, ATF7IP, TREML2, NFKB1, VAMP8, SLC16A3, KIF2A, LMO2, CAPZA2, TBC1D1, GIMAP8, RASSF3, IFNGR2, TONSL, CLEC2D, PTEN, CD226, HLA-DQB1, HLA-DQB2, SGK1, LCN2, RGS14, PTP4A2, TMEM156, MCTP1, ARF1, N4BP1, ARHGAP9, BIRC3, TXK, NPTN, PSME2, ARL6IP5, PSTPIP1, TUBA4A, SERTAD1, DIAPH1, TNFRSF13B, BATF, IL10RB, ARL5C, CCDC88C, CDK9, SH3BP1, GIMAP4, RNF144A, BCL11A, UBE2D2, RASSF2, ARPC3, C19orf53, GRAMD1A, NADK, TNFRSF13C, TRA2A, GGNBP2, ISG15, DEF6, STAT4, ATP6V0E1, PRRC2C, CARD19, CLEC12A, GSAP, MFNG, CMTM6, GPR65, DYNLT2B, CNTRL, S100A11, TACC1, TMEM59, ZNF292, SH3KBP1, GPR18, SYF2, POLR1F, RINL, GP9, AIM2, CTCF, CD300E, DCK, OTULIN, PIP4K2A, MAX, TNFSF14, CALM1, KRIT1, MYH9, TMPO, TNF, UBL3, CSNK1A1, CEACAM7, CEACEM5, CEACAM6, CEACAM3, CEACAM1, CEACAM8, BMX, YY1, VRK1, PRKCB, PFKFB4, G3BP2, ABTB1, EPSTI1, RIN3, ITPKB, MYL6, MS4A6A, MS4A6E, LY9, TAP2, PRKAR2B, SF3B6, FAM78A, ABHD8, ZDHHC18, ATP1A3, PPP1R10, MCMBP, TWF2, KRAS, ARHGAP4, TMEM71, ALCAM, NOTCH1, MORC3, SUN2, ALDOA, HAUS8, RSRC2, CDC42EP3, IKBKB, CHD7, ARL11, UBB, CLEC6A, CXCR4, STK24, NFKBIZ, SERINC3, TIMM10B, CTSW, RCHY1, TAP1, MAF1, RBM5, SEMA4A, MAT2B, CYTH1, NFKBID,WSGR Docket No. 62193-701.601 CD4, GPI, MEF2A, MARCHF2, JUN, GRAP2, AVL9, FCER2, HBA1, HBA2, BCL2A1, ARF6, RNF13, BLNK, IL27RA, KCNN4, RAC1, FPR1, SASH3, FIS1, ATP8B4, SFXN5, GPR141, RSU1, PRR5L, SCNN1A, TMEM243, FCRLA, PIK3R1, NLRP12, MILR1, and any combination thereof.

[0130] In some embodiments, the biomarkers for the subpopulation of maternal cells is Coro1a, Cd52, Cyba, Rac2, Laptm5, or Ptprc. In some embodiments, the biomarkers for the subpopulation of maternal cells is Coro1a, Cd52, Cyba, Rac2, Laptm5, or Ptprc. In some embodiments, the biomarker for the subpopulation of maternal cells is Coro1a. In some embodiments, the biomarker for the subpopulation of maternal cells is Cd52. In some embodiments, the biomarker for the subpopulation of maternal cells is Cyba. In some embodiments, the biomarker for the subpopulation of maternal cells is Rac2. In some embodiments, the biomarker for the subpopulation of maternal cells is Laptm5. In some embodiments, the biomarker for the subpopulation of maternal cells is Ptprc.

[0131] In some embodiments, the one or more maternal cell biomarkers are located in the subpopulation of fetal cells. In some embodiments, the one or more maternal cell biomarkers are located on the subpopulation of fetal cells.

[0132] In some embodiments, the one or more maternal cell biomarker is an extracellular biomarker. In some embodiments, the one or more maternal cell biomarker is an extracellular protein. In some embodiments, the extracellular protein is a cell surface protein. In some embodiments, the cell surface protein is a cell-surface receptor. In some embodiments, the extracellular protein is a soluble cell surface protein. In some embodiments, the extracellular protein is a transmembrane protein. In some embodiments, the extracellular protein is a transmembrane channel protein.

[0133] In some embodiments, the one or more maternal cell biomarkers is an intracellular biomarker. In some embodiments, the one or more maternal cell biomarkers is an intracellular protein. In some embodiments, the intracellular protein is an enzyme. In some embodiments, the intracellular protein is a phosphoprotein. In some embodiments, the intracellular protein is a growth factor. In some embodiments, the intracellular protein is a structural protein. In some embodiments, the one or more maternal cell biomarkers is a nucleic acid molecule. In some embodiments, the nucleic acid molecule is DNA. In some embodiments, the nucleic acid molecule is RNA. In some embodiments, the RNA is mRNA. In some embodiments, the RNA is rRNA. In some embodiments, the RNA is lncRNA. In some embodiments, the RNA is miRNA. In some embodiments, the RNA is siRNA.WSGR Docket No. 62193-701.601

[0134] In some embodiments, the ex vivo population of cells is quantified. In some embodiments, the subpopulation of fetal cells in the ex vivo population of cells is quantified. In some embodiments, the subpopulation of maternal cells in the ex vivo population of cells is quantified. In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells in the ex vivo population of cells is quantified. In some embodiments, the ex vivo population of cells is quantified by flow cytometry. In some embodiments, the ex vivo population of cells is quantified by fluorescence-activated cell sorting (FACS) flow cytometry. In some embodiments, the ex vivo population of cells is isolated through magnetic-activated cell sorting (MACS). In some embodiments, the ex vivo population of cells is quantified by microscopic microdissection. In some embodiments, the ex vivo population of cells is quantified by microscopic imaging. In some embodiments, the ex vivo population of cells is quantified by acoustic focusing flow cytometry. In some embodiments, the ex vivo population of cells is quantified by a hemocytometer. In some embodiments, the ex vivo population of cells is quantified by an automated cell counter. In some embodiments, the ex vivo population of cells is quantified by spectrophotometry. Reagent Compositions

[0135] In some embodiments, the present disclosure provides reagents for use in the meth ods disclosed herein. In some embodiments, the reagents are isolation agents. In some embodiments, the isolation agents couple to one or more biomarkers. In some embodiments, the isolation agents hybridize to the one or more biomarkers. In some embodiments, the isolation agents bind to the one or more biomarkers. In some embodiments, the isolation agents covalently bind to the one or more biomarkers. In some embodiments, the isolation agents bind to the one or more biomarkers via a polar covalent bond. In some embodiments, the isolation agents bind to the one or more biomarkers via a non-polar covalent bond. In some embodiments, the isolation agents non-covalently bind to the one or more biomarkers. In some embodiments, the isolation agents bind to the one or more biomarkers via hydrogen bonding. In some embodiments, the isolation agents bind to the one or more biomarkers via ionic bonding. In some embodiments, the isolation agents bind to the one or more biomarkers via halogen bonding. In some embodiments, the isolation agents bind to the one or more biomarkers via Van der Waals forces. In some embodiments, the isolation agents bind to the one or more biomarkers via electrostatic interactions. In some embodiments, the isolation agents bind to the one or more biomarkers via London dispersion forces. In some embodiments, the isolation agents bind to the one or more biomarkers via hydrophobic effect. In some embodiments, the isolation agents couple to one or more fetal biomarkers. In some embodiments, the isolation agents couple to one or moreWSGR Docket No. 62193-701.601 maternal biomarkers. In some embodiments, the isolation agents couple to one or more fetal biomarkers and one or more maternal biomarkers. In some embodiments, the reagent is a biological agent. In some embodiments, a biological agent is an isolation agent. In some embodiments, the biological agent is an affinity reagent.

[0136] In some embodiments, the biological agent recognizes a fetal biomarker. In some embodiments, the fetal biomarker is located in and / or on a fetal cell. Non-limiting examples of fetal cell biomarkers include H19, 1600025M17Rik, Sct, Ctsr, Rhox9, Prl2c5, Rhox6, Fthl17a, Ctsj, Prl3b1, Nrk, Prl2b1, Ctsq, Hand1, Rhox5, Prl2c2, Krt8, Prl2c3, Prl7a1, Trap1a, Prl3d1, Igf2, Krt18, Nup62cl, Prl7d1, Rhox12, Prl2a1, Krt19, Plac1, Prl3d2, Serpinb9g, Slc38a4, Prl4a1, Serpinb9d, Serpinb9e, Prl8a9, Cited1, Sparc, Gm9112, Ldoc1, Serpinb9f, 3830417A13Rik, Serpinb9c, Tinagl1, Tfpi, Dusp9, Cts6, Hspb1, Nxf7, Sult1e1, Fnd3c2, Rarres2, Procr, Fermt2, Ghrh, Prl2c1, Plet1, Sbsn, Cdh5, Psg19, Igf2bp1, Tpbpa, Cda, Erv3, Pramef12, Shroom1, Peg10, 1700011M02Rik, Psg29, Cyr61, Crct1, Prl8a1, Psg22, Chrdl2, Xlr5a, Prl3d3, Ceacam9, Inhbb, Krt7, Serpinb9b, Cdkn1c, Rbp1, Mrgprg, Gm7120, Aoc1, Pla2g5, Rhox1, Tfap2c, A2m, Tpbpb, Lmcd1, Cryaa, Prl5a1, Zfp36l3, Gm9, Prl6a1, Rgs17, Bex1, Cdh3, Prl7b1, Hsd17b2, Cyp17a1, Gm9513, Tex19.1, Parva, Papl, Map1b, Gpr50, 1600015l10Rik, Ceacam13, Prom2, Slco5a1, Cts7, Nos2, Nppc, Arhgap42, Ppp2r2c, Peg3, Mfap5, Tnfrsf9, Prl8a2, Ceacam15, Rimklb, Aard, Prl7a2, Slc30a2, 4933402E13Rik, Mmp1a, Epn3, Irx3, Fam83b, Tead4, Lama1, Htra1, Pfpl, Cts8, Adgrf5, Prl8a6, Drd4, Bmp1, Cystm1, Gml2, Gm13889, Krtdap, Pappa2, Sema3e, Homer2, Krt14, Pla2g2f, Klhl13, 2310030G06Rik, Car4, Gm7257, Prss8, Dsc2, Ptprn2, Des, Hsd3b6, Pxdn, 270033N17Rik, Eps8l2, 1700001C19Rik, Ceacam11, 1700089L19Rik, Trpm5, 1600012P17Rik, Got1l1, Dmrtc1b, Mt4, Spint1, Mt2, Fscn1, Gm14547, Serpine1, Col4a1, Ndufa4l2, Dst, Pvrl2, Doxl2, Cyp11a1, Klk6, Atf7ip2, Gm805, Pou2f3, Gpr173, Afap1l2, Nkain1, Fabp6, Triml2, Cryab, Pdgfra, Psca, Scnn1b, Rab15, Fmr1nb, Adamts7, Igfbpl1, A4galt, Hrct1, Aif1l, Gm16136, Sema3f, Fam189a1, Psg27, Gm22, Gm26829, Igf2os, Rpl29, Eif2s3y, Il25, Actn3, Sez6l, Unc5d, Frem2, Gm20714, Tff1, Hrc, Hspg2, Pdia5, Ccdc155, 2310016G11Rik, Fam90a1b, Vgf, Cts3, Klk4, Ttc9, Ces1a, Zfp385b, 4631405K08Rik, Nptx2, Adra2b, Mettl24, Gm10251, Gm4758, Prl7c1, Col4a2, Cmtm5, Il24, Ephb3, Cartpt, Foxf1, Gm11755, Spdye4a, Myt1l, 4930562A09Rik, 9930038B18Rik, Msx1, Elf3, Adgrf4, Dio3, Rgs5, Star, Bcat1, Podxl, Lepr, Col5a3, Slco2a1, Myom3, Gm9979, Gpnmb, Fam159a, Trim29, Aqp8, Syt9, Kng2, Wnt7b, St8sia5, Tmem125, Dysf, Uaca, Fetub, Bcar1, Pcdh12, Gm14393, Fabp3, Stra6, Osmr, Lamc2, Muc1, RP23-378L12.3, Bok, Fdx1, Pkp2, Acbd7, Cyb5r3, Sdc1, Sfrp5, Lama5, Slc39a2, Zfp42, Sox3, Vvstm4, Gpx3, Dmkn, Gjb3, Lgi2, Stard10, Slc6a2, Prune2, H19, Sct, Rhox9, Krt18, Prl2c5, Rhox6, Krt8, Prl2c3, Ctsj,WSGR Docket No. 62193-701.601 Krt19, Prl3b1, Prl3d1, Prl7d1, Prl7a1, Tfpi, Trap1a, Fthl17a, Ctsr, Sparc, Igf2, Prl2c2, Hspb1, Nrk, Ctsq, Rhox5, Prl2b1, Rarres2, Hand1, Peg10, Prl2a1, Procr, Sbsn, Prl4a1, Rhox12, Cdkn1c, Ghrh, Cdh5, Cited1, Gm9112, Krt7, Plac1, 1600025M17Rik, Serpinb9d, Cts6, Prl3d2, Serpinb9e, Dusp9, Map1b, Plet1, Nup62cl, Serpinb9g, Fermt2, Slc38a4, Ldoc1, Serpinb9f, 3830417A13Rik, Prl8a1, Hsd17b2, Prl8a9, Psg29, Nxf7, Psg22, Fnd3c2, Serpinb9c, Bex1, 1700011M02Rik, Tpbpa, Prl3d3, Prl2c1, Parva, Tinagl1, Col4a1, Prl5a1, Adgrf5, Igf2bp1, Rbp1, Mt2, Serpinh1, Tfap2c, Serpinb9b, Klhl13, Prl8a2, Lepr, Sult1e1, Cyp11a1, Gjb3, Gm7120, Htra1, Cts7, Cyp17a1, Cystm1, Pramef12, Tnfrsf9, Hsd3b6, Crip2, Ceacam9, Lgals1, Ctsl, Cryab, Homer2, Slc30a2, Cdh3, Zfp36l3, Dmkn, Nppc, Ceacam15, Syngr1, Shroom1, Fscn1, Hspg2, Fmr1nb, Mmp1a, Sdc1, Pfpl, Krt14, Tex19.1, Psg19, Cb5r3, Sema3e, Peg3, Pdia5, Chrdl2, Dst, Dbi, Meis2, Sema3f, Rgs17, Col4a2, Fdx1, Dmrtc1b, Slco2a1, Tpm1, Stard10, Mdk, Irx3, Maged1, Star, Csrp1, Nbl1, Rnd2, Prdx4, Des, Alpl, Xlr5a, Afao1l2, Aprt, Arhgap42, Vgf, Erv3, Tbpbp, Cst8, P4hb, Tbrg1, Cyr61, Muc1, Tmem37, Podxl, Fabp3, Fkbp9, Fstl3, Car4, Epas1, Tpm2, Rhox1, Chchd10, Stra6, Egfl7, Rab15, Copz2, Nos2, Lgals3, Sin3b, Slc6a2, Epcam, Pkp2, Pla2g5, Pvrl2, Rhox10, Prl7b1, Ada, Lmcd1, Erdr1, Selm, 1700086L19Rik, Dmrtc1a, Bok, Ldhb, Epn3, Mrgprg, Basp1, Gm9, Aoc1, Serpinb6b, Uaca, 2310030G06Rik, Ckb, Cstb, Ssr4, Foxo4, Ngef, Plpp1, A2m, Dstn, Cox6a1, Gale, Pde10a, Krtdap, Rab6b, Esam, Crct1, Col5a2, A4galt, Dnase1l3, Sfn, Bmp1, Calr, Papl, Kank1, Prl6a1, Cmtm5, Ildr2, Rhoc, Ccl27a, Efs, Hrct1, Dsc2, Bcar1, Cd63, Inhbb, Ppp2r2c, Mpdz, Mt1, Fetub, Nid1, Bicc1, Psg23, Hdlbp, Rasd1, Arhgap22, Maged2, Aard, Prl8a6, Cald1, Prom2, Pdia6, Nostrin, Prss8, 1600012P17Rik, Dsp, Adm, Fam90a1b, Exoc3l4, Rbfox2, Mgst3, Gm648, Rgs5, Bgn, Galk1, Adra2b, AT036118, Nenf, Aif1l, Nfib, Acvr2b, Htatsf1, Rpn1, Psg27, Phldb2, Myof, Fbxl19, H1f0, Cobl, Emp2, Myrf, Adamts7, Fam89a, Gm9513, Tmem98, Mfsd2a, Mgat4b, Cav1, Il33, C77080, Lars2, Gm21887, 9930038B18Rik, Prom1, Ptov1, Ltbp4, Mmp23, Prl7a2, Cts3, Fabp6, Prkcdbp, Lmna, Rpn2, Caskin1, Lama1, Fam83b, Cryaa, Lamc2, 4933402E13Rik, Fn1, Dlc1, Lrrn4, Tff1, Elf3, Serpine1, Bcat1, Perp, Lama5, Gpx3, Car2, Pawr, Phactr1, Oaf, Lamb2, Prnp, Scin, Parm1, Smim1, Jup, Gpx8, Atf5, Ift43, Gkap1, H1fx, Cenpt, Rhou, Rrm2, Cul7, Asns, Tubb3, 2200002D01Rik, Abhd6, Hdac6, Rgs16, Cgnl1, Atp9a, Src, 4930486L24Rik, Elf3, Ly6c1, transferrin receptor (TfR), ZNF614, ZNF539, ZNF283, ZFYVE9, WRN, UBL4A, TPM3, TP73, TMEFF2, THC2274391, THC2265980, t (BRACHYURY), SYT9, SLAMF1, RPS27, RPL39, RPL26, RPL23A, RPL23, RP11-78J21.1, RGPD2, QPRT, PPIA, PGK1, PF4, NTRK1, NBR2, MYL6, MAPK3, MAD2L1, LOC389286, KY, KMO, KCNK4, ITGA7, HMGN2, HMGA1, HLA-C, HIST1H2AJ, GSK3A, GCC2, FYN, F1135740, ENST00000356196, EEF1A1, EDN1, DKFZP434F142, D4ST1, CRYL1, CPS1,WSGR Docket No. 62193-701.601 CD7, CCNA2, CB123670, BC111482, 6C089454, ALS2CL, AK000420, AHNAK, ADAM11, ACVR2B, ABHD2, CD105, HLA-G, galectin 13 (LGAGLS13 / PP13), galectin 14 (LGALS14), placental growth factor (PGF), pregnancy-associated plasma protein A (PAPPA), alpha fetal protein (AFP), endoglin (ENG), fns-related tyrosine kinase 1 (FLT-1), keratin-7, PLAP, MCAM, laeverin, H315, FT1.41.1, NDOG-1, NDOG-5, BC1, AB-154, AB-340 (PAR-1), Glut- 12, factor XIII, hPLH, HLA-C, JunD, Fra2, NDPK-A, CAR, HASH2, αHCG, IGF-11, PAI-1, p57 (KIP2), PP5, PLAC1, PLAC8 and PLAC9, CHL1, H119, HLA-C, Fra2, NDPK-A, the CAR protein, HASH2, human chorion gonadotropin alpha (hCGα), TFPI-2, annexin IV, cytokeratin- 7, cytokeratin-8, cytokeratin-19, H19, IFG2, ESX1L, MASH2, ASH2, Stra13, FosB, Cyclin D1, GCM1, caspase-8, factor XIII, tapasin, Connexin 31, Connexin 43, HAND1, Syncytin, MMP9, APAF-1, caspase-3, caspase-9, FAS, Fas ligand, FLIP, AP-2γ, 313-HSD VI, CDX2, ERR2, PLX3, PTHrP, ASCL2, ID2, MET, TEF5, UPA, 11β-HSD2, c-ETs1, HMGI(Y), estrogen receptor, GemI, TTPA, 1503-7E, J42-4d, J2r(3), J2r(12), J2r(13), 305-4G, K1-1a, K2r / 1f(50), K2r / 1f(59), K(1)157-2A, K3r(HIGH)76, 597-10C, NT7-T3, N9r.Mf, 334-2C, O19r-T3, O1-1a, 332-9E, P60-1a, P1-1a, P3r(9), 305-9E, R5’-T3, R6r / 1-6H, 369-8G, U2f-T3, 305-6G, L15-1a, L21-1a, 252, 120r, clone-1, D19-2g, CD146, CD141, vimentin, VCAM, ICAM, VEGFR-1, VEGFR-2, VEGFR-3, ITGA5, ITGB5, CDH11, CDH3, CK1, CK2, CK3, CK4, CK5, CK6, CK7, CK8, CK9, CK10, CK13, CK14, CK15, CK16, CK17, CK18, CK19, CD9, CD39, CD200, EPHB4, EPCR, PAR-1, hPL, CSH2, KISS1, GDF15, CRH, TFPI2, β-hCG, LOC90625, FN1, COL1A2, PSG9, HBE, AFP, GC, APOC3, SERPINC1, APOB, AHSG, HPX, CPB2, ITIH1, APOH, AMBP, Thy-1, CD90, thromobomodulin, Flt-1, EPCR, CDH11, CDH3, CD59, CD71, CD36, Glycophorin A, hexokinase 2, placental alkaline phosphatase, hyalouronic acid, BCL-2, VE-cadherin, PECAM-1, α1-integrin, placental lactogen, PSG-1, α6 integrin, E-cadherin, MMP14, KCNQ4, CLDN6, F3, EPCAM, TROP2, CD147, fetal hemoglobin, embryonic hemoglobin, ITGA6, FSHR, CD45, LHCGR, CD270, Trop2, FOLR1, CD114, CSF3R, SLC40A1, CD156a, ERVW1, GPC3, TREML2, ALPP (866), CD163, CD144, ALPP (GM022), FOLR1, CD115, TSPAN1, SLC22A1, EFNA1, SLC62A2, SLC40A1, CD71, CD166, TGF- beta2, human placental lactogen, c-erbB2, PAPPA2, PRG2, cytokeratin 7, beta-HCG, ,alpha- HCG, GCM1, syncytin, c-erbB2, leptin, INSL4, TGF-beta1, CSH1, KISS1, PEG10, PAGE4, p63, miR1246, miR1323, miR-512-3p, has-miR-516b-5p, miR-517-5p, has-miR525, miR-526b, ath-miR159a, RNU44, RNU48, U6, CDH9, HLA-G, MIH61, ALPP (8B6), CD49e, JAM2, transferrin receptor, glycophorin a (GPA), EGFR, thrombospondin receptor (CD36), CD34, HbF, HAE 9, FB3-2, H3-3, erythropoietin receptor, HBE, AFP, APOC3, SERPINC1, AMBP, CPB2, ITIH1, APOH, HPX, AHSG, APOB, J42-4-d, 2,3-biophosphoglycerate (BPG), carbonicWSGR Docket No. 62193-701.601 anhydrase (CA), thymidine kinase (TK), MMP14, lectin, CD235a, carbohydrates, selectin, GPA, ANTIGEN-I, EpCAM, E-cadhrin, Muc-1, Hpl, CHS2, KISS1, GDF15, CRH, TFP12, CGB, LOC90625, FN-1, COL1A2, PSG9, PSG1, EGFR, CD105, cytokeratin, TLS1, CD147, CD147, PTPRN, BMPR2, TNFRSF19, STIM1, SELPLG, KLP-1, HECA452, GPR160, KLRD1, TNFSF18, LRP11, ABCA1, GPR176, SCARF, ESYT1, SCARB1, LTB4R, TLT2, CD141, CD90, Vimentin (Vim), VCAM, ICAM-1, VEGFR-1, Flt-1, VEGFR-1, VEGFR-2, VEGFR-3, PAI-1, EPCR, CD146, ITGA5, ITGB5, CDH11, CDH3, CD59, CSH-1, CSH-2, PSG2, PSG3, PSG11, HBG2, XAGE3, CSHL1, HBG1, ERVH48, phCG, any human orthologue of any of the foregoing, and any combination thereof.

[0137] In some embodiments, the fetal biomarkers of the present disclosure have human orthologues. Non-limiting examples of human orthologues of the fetal biomarkers include: AOC1, FAM236B, FAM236D, FAM236A, FAM236C, C11orf52, MAGEA10, A2M, A4GALT, AARD, ACVR2B, ADA, ADAMTS7, ADGRF5, ADM, ADRA2B, AFAP1L2, AIF1L, ALPL, APRT, ARHGAP22, ARHGAP42, BASP1, BCAR1, BEX1, BEX2, BGN, BICC1, BMP1, BOK, BSG, KIAA1522, CALD1, CALR, CA4, CASKIN1, CAV1, CCL27, CD63, CDA, CDH3, CDH5, CDKN1C, CEACAM4, PSG3, PSG8, PSG1, PSG6, PSG7, PSG11, PSG2, PSG5, PSG4, PSG9, CHCHD10, CHRDL2, CITED1, CKB, CMTM5, COBL, COL4A1, COL4A2, COL5A2, COPZ2, COX6A1, CRIP2, CRYAA, CRYAB, CSRP1, CSTB, CYB5R3, CYP11A1, CYP17A1, CCN1, CYSTM1, DB1, DCN, DES, DLC1, DMKN, DNASE1L3, DSC2, DSP, DST, DSTN, DUSP9, EFS, EGFL7, EMP2, EPAS1, EPCAM, EPN3, EPS8L2, ESAM, EXOC3L4, FABP3, FABP6, FAM83B, FAM89A, FAM90A1, FAM90A26, FAM90A7, FAM90A22, FAM90A23, FAM90A14, FAM90A18, FAM90A16, FAM90A8, FAM90A17, FAM90A19, FAM90A9, FAM90A10, FBXL19, FDX1, FERMT2, FETUB, FKBP9, FMR1NB, FN1, FOXO4, FSCN1, FSTL3, GALE, GALK1, GHRH, GJB3, CT45A1, CT45A3, CT45A5, CT45A6, CT45A2, CT45A7, CT45A8, CT45A9, CT45A10, SAGE1, TMEM267, GPR50, H1- 0, HAND1, HDLBP, HMGN1, HOMER2, HRCT1, HSD17B2, HSPB1, HSPG2, HTATSF1, HTRA1, IGF2, IGF2BP1, IL33, ILDR2, INHBB, IRX3, KANK1, KLHL13, KRT18, KRT19, KRT7, KRT8, KRTDAP, LAMA1, LAMC2, LARS2, LDHB, LSOC1, LEPR, LGALS1, LGALS3, LMCD1, LMNA, LRRN4, LTBP4, MAGED1, MAGED2, MAP1B, MDK, MEIS2, MFSD2A, MGAT4B, MGST3, MMP1, MMP23B, MPDZ, MRGPRG, MT1G, MT2A, MUC1, MYOF, MYRF, NBL1, NENF, NFIB, NGEF, NID1, NOS2, NOSTRIN, NPC2, NPPC, NRK, NUP62CL, NXF5, NXF2, NXF2B, P4HB, ACP7, PARVA, PED10A, PDIA5, PDIA6, PEG10, PEG3, MPEG1, PHLDB2, PKP2, PLA2G5, PLAC1, PLET1, PLPP1, PODXL, PPIB, PPP2R2C, PRAMEF12, PRAMEF1, PRAMEF11, PRAMEF2, PRAMEF4, PRAMEF10,WSGR Docket No. 62193-701.601 PRAMEF7, PRAMEF6, PRAMEF27, PRAMEF25, PRAMEF26, PRAMEF9, PRAMEF13, PRAMEF18, PRAMEF5, PRAMEF8, PRAMEF33, PRAMEF 15, PRAMEF14, PRAMEF19, PRAMEF17, PRAMEF20, PRDX4, PRKCD, PROCR, PROM1, PROM2, PRSS8, CEACAM4, PTOV1, NECTIN2, RAB15, RAB6B, RARRES2, RASD1, RBFOX2, RBP1, RGS17, RGS5, RHOC, RND2, RPN1, RPN2, S100A6, SBSN, SCT, SDC1, SEC61G, SEMA3E, SEMA3F, SERBINB6, SERBINB9, SERPINH1, SFN, SHROOM1, SIN3B, SLC30A2, SLC38A4, SLC6A2, SLCO2A1, SPARC, SSR4, STAR, STARD10, STRA6, SULT1E1, SYNGR1, TBRG1, TEX19, TFAP2C, TFF1, TFPI, TINAGL1, TMEM37, TMEM98, TNFRSF9, TPM1, TPM2, UACA, UQCRB, ATP5MK, VGF, FAM9A, FAM9B, FAM6C, C19orf33, ABHD6, ASNS, ATF5, ATP9A, BCAT1, CA2, CENPT, CGNL1, CTSL, CUL7, ELF3, FBXL19, GKAP1, GPX3, GPX8, H1FX, HDAC6, IFT43, JUP, LAMA5, LAMB2, OAF, PARM1, PAWR, PERP, PFPL, PHACTR1, PRNP, RGS16, RHOU, RRM2, SCIN, SERPINB6, SERPINB9, SERPINE1, SMIM1, SRC, TUBB3, and any combination thereof .

[0138] In some embodiments, the biological agent recognizes a Sct biomarker or a human orthologue of a Sct biomarker. In some embodiments, the biological agent recognizes a H19 biomarker or a human orthologue of a H19 biomarker. In some embodiments, the biological agent recognizes a Tfpi biomarker or a human orthologue of a Tfpi biomarker. In some embodiments, the biological agent recognizes a Fthl17a biomarker or a human orthologue of a Fthl17a biomarker.

[0139] In some embodiments, the biological agent recognizes a maternal biomarker. In some embodiments, the maternal biomarker is located in a maternal cell. In some embodiments, the maternal biomarker is located on a maternal cell. Non-limiting examples of maternal cell biomarkers include Coro1a, Rac2, Malat1, Laptm5, H2-D1, B2m, Cytip, Alox5ap, Cd52, Junb, Cyba, Fyb, Btg1, Srgn, Actb, Klf2, Mcl1, H3f3a, Hmha1, Tmsb4x, Cd53, Ptprc, Fxyd5, Myl12b, Tyrobp, Gmfg, Fcer1g, Ier2, H2-K1, Itm2b, Jund, Rasgrp2, Nfkbia, Itgb2, Pfn1, Ifngr1, Ltb, Pim1, Ssh2, Rbm39, Sell, Trbc2, Msn, Ddx5, Stk17b, Lyz2, Cd37, Csf3r, Hcst, Ifitm2, Pnrc1, Rap1b, Meat1, Fth1, Cd3d, Arhgdib, Pglyrp1, H3f3b, Ms4a6b, Akap13, S100a9, Lyn, Sorl1, Mbnl1, Cd44, Ier5, Lsp1, Cdc42, Aes, Ms4a6c, Msrb1, Gdpd3, Btg2, Sat1, Arhgap30, Lrg1, Itga4, Oaz1, Rhoa, Eif1, Zfp36l2, Selplg, Cd3g, Zfp36, Arhgap15, Syk, Arpc2, Sf3b1, Ighm, Psmb9, Gnai2, Actr3, Lst1, Il1b, Satb1, Amp32a, Foxp1, Lcp1, Wfdc17 , Taldo1, Slfn2, Rgs10, Ccnd3, Ncor1, Cd3e, Vim, Cebpb, Tpm3, Adgre5, S100a8, Ccl6, Ptpn18, Napsa, Rps9, C5ar1, Hnrnpf, Iqgap1, Ly6e, Klf13, Ifitm3, Psmb8, H2-Q4, Gsr, Hmgb2, Fos, Sh3bgrl3, Dusp1, Spi1, Cd84, Tspo, Lef1, Cyp4f18, Atp11b, Ccr1, Gpx1, Cd47, Samhd1, AB124611, Trac, AI467606, Shisa5, Myl6, Tspan13, Cfl1, Gm5150, Itgal, Calm2, Plek, Ucp2, Tsc22d3,WSGR Docket No. 62193-701.601 Ikzf1, Grk6, Gimap1, Ybx1, Gm8797, Clic1, Rara, Limd2, Ifi27l2a, Ubb, Gngt2, Cd14, Pla2g7, Ets1, Tgfb1, Actg1, Il17ra, Celf2, Srsf5, Gimap6, Tmem50a, Ctss, Ube2d3, Tln1, Apbb1ip, Gabarap, Arpc5, Kmt2e, Klf3, Cap1, Itgam, Emp3, Clk1, Ogt, Crlf3, Samsn1, Stk38, Tax1bp1, Lat, Clec4e, Cdkn2d, Rel, Srrm2, Smc6, Morf4l1, Rsrp1, Ptpn1, Rgs2, Fmnl1, Gpsm3, Klf6, Fcgr3, Mef2c, Birc2, Kdm6b, Tra2b, Ptprcap, Son, Rassf5, Rpl22, Gimap3, Hnrnpa2b1, Lbr, Lck, Calm1, Cdkn1b, Ccr7, Ypel3, Ptpn6, Cd74, Ppp2r5a, Cd9, Rcsd1, Svil, Hmgb1, Rhog, Hacd4, 9930111J21Rik2, Stat1, Ncf4, Fam105a, Rsbn1l, Hp, Ankrd11, Emb, Dusp2, Pfdn5, Txnip, Psme1, Slfn1, Ostf1, Jak1, Sirpa, Rap1a, Fam49b, Gda, Picalm, Ywhaz, Pde4b, Dazap2, Tsc22d4, Pkm, Slc25a5, Kdm7a, Lcp2, Fus, H2-Q7, Mxd1, Glipr1, Bnip3l, Luc7l2, Tnfaip8l2, Pdcd4, Map1lc3b, Plbd1, Skap1, Unc93b1, Ncf2, Rnf149, Bin2, Mycbp2, Cnp, S100a10, Ubc, Cd300ld, Fam32a, Ptp4a2, Adrb2, Prdx5, Crip1, Sna32, Csk, Tnrc6b, Arhgef1, Pou2f2, Hspa8, Pilra, Bcl11b, Zfp292, Cnn2, Aldh2, Ywhah, Zbtb7a, Pycard, Cotl1, Retnlg, Sp100, Irf2, Fgr, Igsf6, Ablim1, Fam65b, Fermt3, Tmbim6, Tpd52, Scp2, Ube2b, Notch2, Gnb2, Il7r, Capzb, Jun, Ptma, 2810474O19Rik, Prr13, Mrpl33, Ppp1r15a, Lilrb4a, Zyx, St8sia4, Arpc3, H2afz, S100a11, Pag1, Irf1, Tm6sf1, Rsf1, Frina, Rasa3, Smap2, Dok3, Skap2, Add3, Stx11, Mkrn1, Hck, Ccnl1, Ppp1ca, R3hdm4, Cd48, Chd2, Rgs19, Cth4, Atp2b1, Peli1, Fam107b, Ptbp3, Adipor1, Serp1, Pak2, Hspa5, Tpr, Zc3hav1, Sdcbp, Dock2, Rnf130, Ftl1, B4galnt1, Cd300a, Hcls1, Anxa1, Cst3, Ankrd44, Elf1, Prpf38b, Atp6v1g1, Snx20, Il6ra, Whsc1l1, Tmem71, Stk4, Sla, Mbp, Dnaja1, Vgll4, Capza2, Ldha, Pyhin1, Rapgef6, Hdc, Fli1, Rbm25, H2afy, Prdx6, Rac1, Stap1, Supt4a, Hnrnpk, Ubl5, Cdk2ap2, Ppp1r2, Psenen, Bcl2, Tprgl, Arpc1b, Cd69, Phf20l1, Ppt1, Rhoh, Coro1a, Cytip, Malat1, Rac2, Cd52, Laptm5, B2m, H2-D1, Cyba, Ptprc, Btg1, Gmfg, Junb, Srgn, Fxyd5, Itgb2, Ltb, Tyrobp, Mcl1, Cd53, Klf2, Fcer1g, H3f3a, Hmha1, Itga4, Alox5ap, Ptpn18, Myl12b, Cd3d, Spi1, Ier2, Ccl6, Ms4a6b, Msn, Selplg, Pglyrp1, S100a9, Lyn, Slfn2, H2-K1, Ier5, Cyp4f18, Csf3r, Hp, Lst1, Jund, Fyb, Rap1b, Pim1, Il1b, Lyz2, Stk17b, Cd3e, Gpsm3, Lef1, Sell, Hcst, Gm5150, Ms4a6c, Fmnl1, Adgre5, Arhgap30, Lrg1, 9930111J21Rik2, Unc93b1, Rbm39, Pnrc1, Ptpn6, Trac, Akap13, Actr3, Ifitm2, Crlf3, Msrb1, Samsn1, Ets1, Lck, Pla2g7, Zfp36l2, Lat, Ncor1, Sorl1, Ccnd3, Aes, Rasgrp2, Samhd1, Psmb8, S100a8, Wfdc17, Ctss, Neat1, Arhgap15, Ncf4, Lcp1, AB124611, Ssh2, Btg2, March1, Snx20, Fam105a, Zfp36, C5ar1, Tspan13, Hck, Apbb1ip, Mbnl1, Cd37, Limd2, Skap1, Iqgap1, Cybb, Lcp2, Ly6e, Slfn1, Bcl11b, Syk, Sf3b1, Ccr1, Ddx5, Stk38, Sat1, Anp32a, Dock2, Cd74, Satb1, Ighm, Foxp1, Tspo, Klf13, Ptpn1, AI467606, Nfam1, Cap1, Hnrnpf, Shisa5, Fgr, Rhog, Ifngr1, Psmb9, Rhoh, Lilrb4a, Plek, Rasa3, Sla, Atp11b, Fam65b, Gimap1, Hmgb2, Ypel3, Tnfaip8l2, Pirb, Rgs2, Tmem50a, Fcgr3, Cd84, Kmt2e, Mpeg1, Retnlg, Ikzf1, Il6ra, Prdx5, Ostf1, Cd14, Itgam, Cd300ld, Rnf149, Cd79b, Dusp1, Tsc22d4, Ifi27l2a, Il17ra, Sirpb1c, Klf3,WSGR Docket No. 62193-701.601 Celf2, Clic1, Cebpb, Grk6, Pira2, Grap, Ppp2r5a, Fam49b, Cd47, Tgfb1, Wfdc21, Hcar2, Ncf2, Cd300a, Ms4a4b, Itgal, Trbc2, Dusp2, Tra2b, Pilra, Cyth4, Hcls1, Pyhin1, Son, Tgfbi, Tspan32, Vav1, Cd3g, Irf1, Dok3, Cxcr2, Ifitm3, Rara, 2810474O19Rik, Cd33, Adrb2, Snap23, Fos, B4galnt1, Lilr4b, Tpm3, Prkcq, Cd44, Rcsd1, Sp100, S1pr1, Cdk2ap2, Cd247, Cotl1, Lrrfip1, Slc11a1, Srsf5, Cnp, Klf6, Clec4e, Inpp5d, Glipr1, Lpcat2, Fermt3, Clec4a3, Fus, Itgb7, Tbc1d10c, Napsa, Pld4, Add3, Jak1, Aldh2, Cdkn1b, Gsr, Myo1f, Ccr7, Mxd1, St8sia4, Gimap3, Arpc5, Ly6d, Rassf5, Ifitm1, Arhgef1, Gdpd3, Ifitm6, Smap2, Fam107b, Cd224, Mcemp1, Bin2, Hdc, Fam32a, Pou2f2, Kdm6b, Cdkn2d, Vim F630028O10Rik, H2-Q4, Pag1, Rnf130, Smpdl3a, Tpd52, Tmcc1, Grina, Fgl2, Csk, Trbc1, Cd27, D1Ertd622e, Rasal3, Gabarap, Cd28, Ipcef1, Rgs19, Bcl2, 4930523C07Rik, Slc25a5, Rapgef6, Ppt1, Gngt2, Irf2, Psme1, Clec4d, Slc15a3, Pilrb2, Andrk11, Peli1, Tsc22d3, Bank1, Iglc2, Il7r, Stfa2l1, Stat1, Igsf6, Tax1bp1, Nedd9, Tm6sf1, Ly86, Tprgl, 2310001H17Rik, H2-DMa, H2-DMb2, Sirpb1b, Srrm2, Stk4, H2-T23, Abcg1, Klhl6, Ablim1, Evi2a, Bnip3l, Arhgap25, Lrrc25, Ogfrl1, Emp3, Clk1, Mkrn1, Ggnt2, Hacd4, Picalm, Adipor1, Man2b1, Cd48, Lyl1, Sp140, Sephs2, Lbr, Cd69, Phf20l1, Scp2, Ms4a1, Cdc42se1, Rgs10, Sirpa, Stap1, Ddx6, Ppp1r2, Abi3, Acap1, Csf2rb, Tnfrsf18, Alox12, Ebf1, Luc7l2, Flna, Ikzf3, Sept1, Arid4a, Pi16, Gimap6, Dazap2, Ppp1r16b, Lyst, Adrbk1, Nlrp3, Ptprcap, Pycard, Themis2, Lbh, Mycbp2, Rel, Apobec1, Cd2, Chd2, AF251705, Trem1, Dennd1c, Ram189b, Ncf1, Tnfaip2, Pde4b, Ccnl1, Fcgr4, Skap2, Elf1, Il2rg, Slc7a11, Trim30b, Smchd1, H2-Q7, Rasgrp1, Ptpn22, Mrgpra2b, Ptpn22, Mrgpra2b, Cmip, Dgka, 1600010M07Rik, Igkc, Slc2a6, Atg3, Arl4c, Il16, Btk, Il21r, Fli1, Gda, Mbd2, Fam134b, Myo1g, Pygl, Ptpre, Kdm7a, Ctse, Plbd1, Map3k3, Akna, Mmp8, Ywhaz, H2afy, Svil, Mfsd6, Emilin2, Hnrnpa0, Psenen, Pdcd4, Cd5, Chil1, Pnpla2, Macf1, Gm8369, Nfatc1, Plcg2, Rhof, Gm9733, Map1lc3b, Capzb, Ube2b, Ccpg1, Il1f9, Xist, Gimap5, Rsbn1l, Cbl, Cd300lb, Siglece, Slco3a1, Supt4a, Mbp, Ms4a4c, Cd79a, Nrros, Gimap7, Birc2, Ccdc12, Mapk14, Tcf7, Whsc1l1, I830077J02Rik, Rbm25, Xrn2, Jakmip1, Dock10, Vsir, Siglecg, Ramp1, Cd6, Phip, Ifi203, Tpr, Kpna4, Ptafr, Sh3bp5, Itk, Tnrc6c, Notch2, Myd88, Nipbl, Prkcd, Slc2a3, Bcl10, Il10ra, Dhrs7, Clec4a1, S1pr4, Cd300lf, Ogt, Lrmp, Ttc7, Gm43603, Treml1, Bcl2a1b, Marcks, Mier1, Vasp, Clec4a2, Gpcpd1, Traf3ip3, Prpf38b, Runx3, R3hdm4, Lfng, 1110008F13Rik, Slamf6, Pdcd10, Stx11, Srsf11, Gp5, Hnrnpl, BC028528, Gm26740, Pid1, Nfe2, Mzb1, Lilra6, Snca, Parp14, Vgll4, Socs3, Sh2d1a, Dapl1, Gpr183, AW011738, Hbb-bs, Lmnb1, Strbp, H2- Aa, Il18rap, Hipk1, Stag2, Orai1, Smc6, Stk10, Fam111, Treml4, Ctage5, Ebi3, Ap1s2, Neurl3, Trib2, Zc3hav1, Baz1a, Hectd1, Sec11c, Tnrc6b, Amica1, Prkd2, Rnase6, Evl, Vav3, Rab27a, Trim30a, Adam8, Phf3, Gpr132, Sirt7, Tcp11l2, Zcchc7, Itpr2, Bptf, Ace, Icam2, Slfn4, Fcmr, Prkar1a, Asap1, Clec2i, Prr13, Mef2c, Kansl1, Slc40a1, Vps37b, Sdcbp, Cd8b1, Phf11b, Zeb2,WSGR Docket No. 62193-701.601 Sfpq, Ccl3, Anxa1, Pik3cd, Serp1, Arrb2, Ccdc88b, Ctsc, Atf7ip, Treml2, Nfkb1, Vamp8, Slc16a3, Kif2a, Lmo2, Capza2, Tbc1d1, Gimap8, Rassf3, Ifngr2, Ltb4r1, Trem3, Pf4, Clec2d, Pten, Cd226, H2-Ab1, Sgk1, Zcchc11, Lcn2, Rgs14, Ptp4a2, Tmem156, Mctp1, H2-Q6, N4bp1, Arhgap9, Birc3, Txk, Nptn, Psme2, Arl6ip5, Pstpip1, Tuba4a, Sertad1, Diaph1, Tnfrsf13b, Batf, AW112010, Il10rb, Arl5c, Cxcl2, Ccdc88c, Cdk9, Sh3bp1, Gimap4, Rnf144a, Bcl11a, Rassf2, Gramd1a, Nadk, Tnfrsf13c, Tra2a, Ggnbp2, Isg15, Def6, Stat4, Prrc2c, Gm15987, Card19, Clec12a, Gsap, Mfng, BC021614, Cmtm6, Cpr65, Tctex1d2, Cntrl, Tacc1, Zfp292, Sh3kbp1, Ifi47, Gpr18, Syf2, Twistnb, Rinl, Gp9, Aim2, Ctcf, Cd300e, Dck, B430306N03Rik, Otulin, Pip4k2a, Max, Tnfsf14, Krit1, Tmpo, Tnf, Ubl3, Csnk1a1, Fpr2, Cecam1, Bmx, Vrk1, Prkcb, Pfkfb4, G3bp2, Abtb1, Epsti1, Rin3, Itpkb, Gimap9, Adgre4, Ms4a6d, Ly9, Tap2, Prkar2b, Sf3b6, Fam78a, Adhb8, 1700097N02Rik, Zdhhc18, Atp1a3, Ppp1r10, Ifi30, Mcmbp, Twf2, C130050O18Rik, Kras, Arhgap4, Tmem71, Alcam, Notch1, Morc3, Sun2, Haus8, Rsrc2, Cdc42ep3, Ikbkb, Chd7, Arl11, Clec4n, Cxcr4, Stk24, Nfkbiz, Serinc3, Timm10b, Ctsw, Rchy1, Tap1, Maf1, Rbm5, Sema4a, Mat2b, Cth1, Nfkbid, Cd4, Ly6c2, Mef2a, March2, Grap2, Avl9, Fcer2a, Cd24a, Hba-a1, Bcl2a1a, Arf6, Rnf13, Blnk, Il27ra, Kcnn4, Hn1, Fpr1, Sash3, Atp8b4, Sfxn5, Gpr141, Rsu1, Prr5l, Scnn1a, Tmem243, Fcrl1, Pik3r1, Nlrp12, Milr1, THBD, F3, FLT3, MCAM, CD141, CD135, CD142 / F3, CD146, CD33, CD68, CD144, CDH5, CD62E, CD45, CD90, SELE, GAP / GLA5, ITGA5, KCNK3 / TASK-1, CNTFR, JAM-1 (f11r), SDC4 / Syndecan 4, ACVR2B, ACVR2A, CD44v6+CD44v4 / 5, STX1A, TEK / Tie2, CD171, L1CAM, NCAM1 / CD56, ALPP, ALPPL2, MMP23A / MMP23B, CD14, CD4, CD8, CD3, CD19, CD32, CD16, HLA-A, HLA-B, CD15, FSHR, CD45, LHCGR, CD270, Trop2, FOLR1, CD114, CSF3R, SLC40A1, SLC22A1, SLC22A1 CD156a, ERVW1, GPC3, TREML2, ALPP (866), CD163, CD144, ALPP (GM022), FOLR1, CD115, TSPAN1, SLC22A1, EFNA1, SLC62A2, SLC40A1, CD71, CD166, CDH12, HLA-A, HLA-B, PRSS8, CD45, CD230, ALPP (8B6), CD49e, JAM2, TREML2, MMP14, lectin, CD47, CD45, CD35, CD12, CD14, CD32, CD235a, carbohydrates, selectin, GPA, ANTIGEN-I, EpCAM, E-cadherin, Muc-1, Hpl, CHS2, KISS1, GDF15, CRH, TFP12, CGB, LOC90625, FN-1, COL1A2, PSG9, PSG1, CD105, TLS1, CD147, CD15, TNFRSF19, STIM1, CD19, CD14, any human orthologue of any of the foregoing, and any combination thereof.

[0140] In some embodiments, the maternal biomarkers of the present disclosure have human orthologues. Non-limiting examples of human orthologues of the maternal biomarkers include: CYBA, CORO1A, CYTIP, LAPTM5, CD53, RAC2, CD52, PTPRC, FCER1G, ARHGAP45, GPM6A, MAF, C3, CFH, EFEMP1, BICC1, COL1A2, COL1A1, B2M, BTG1, GMFG, JUNB, SRGN, ITGB2, LTB, TYROBP, MCL1, KLF2, NFKBIA, H3F3A, ITGA4, ALOX5AP,WSGR Docket No. 62193-701.601 PTPN18, CD3D, SPI1, IER2, CCL15-CCL14, CCL14, CCL15, CCL23, MS4A6A, MS4A6E, MSN, SELPLG, ACTB, PGLYRP1, TMSB4Y, LYN, SLFN12, SLFN12L, IER5, CYP4F3, CSF3R, HP, HPR, LST1, ITM2B, JUND, FYB1, LSP1, RAP1B, PFN1, PIM1, IL1B, STK17B, CD3E, GPSM3, LEF1, SELL, HCST, AL049634.2, SIRPB1, SIRPG, SIRPA, MS4A6A, MS4A6E, FMNL1, ADGRE5, ARHGAP30, LRG1, UNC93B1, RBM39, PNRC1, PTPN6, AKAP13, ACTR3, IFITM2, IFITM1, IFITM3, ARHGDIB, CRLF3, MSRB1, SAMSN1, ETS1, LCK, PLA2G7, ZFP36L2, LAT, NCOR1, SORL1, CCND3, AES, RASGRP2, SAMHD1, PSMB8, S100A8, CTSS, ARHGAP15, NCF4, LCP1, C19orf38, SSH2, BTG2, MARCH1, GNAI2, SNX20, FAM105A, ZFP36, C5AR1, TSPAN13, HCK, APBB1IP, MBNL1, CD37, LIMD2, SKAP1, IQGAP1, CYBB, LCP2, EMB, LY6E, SLFN12, SLFN12L, BCL11B, SYK, SF3B1, CCR1, DDX5, STK38, SAT1, ANP32D, ANP32A, TALDO1, CDC42, DOCK2, CD74, SATB1, IGHM, TSPO, KLF13, H3F3B, PTPN1, C16orf54, NFAM1, CAP1, HNRNPF, SHISA5, FGR, RHOG, IFNGR1, PSMB9, RHOH, PLEK, RASA3, SLA, ATP11B, RIPOR2, SH3BGRL3, GIMAP1, HMGB2, RHOA, YPEL3, TNFAIP8L2, LILRB3, LILRA6, LILRB5, LILRA4, ARPC2, RGS2, TMEM50A, FCGR2A, FCGR2C, FCGR2B, CD84, KMT2E, MPEG1, RETNLB, TXNIP, IKZF1, IL6R, PRDX5, OSTF1, CD14, ITGAM, CD300LF, RNF149, CD79B, DUSP1, TSC22D4, IL17RA, AL049634.2, KLF3, CELF2, CLIC1, CEBPB, GPX1, GRK6, LILRA5, GRAP, GRAPL, PPP2R5A, FAM49B, CD47, TGFB1, HCAR2, HCAR3, NCF2, CD300A, CD300LD, ITGAL, TRBC1, TRBC2, OAZ1, DUSP2, TRA2B, CFL1, CYTH4, ANKRD44, HCLS1, SON, TGFBI, TSPAN32, VAV1, CD3G, IRF1, MRPL33, DOK3, CXCR2, IFITM1, IFITM3, IFITM2, RARA, KIAA1551, CD33, SIGLEC6, ADRB2, SNAP23, FOS, B4GALNT1, PRKCQ, CD44, RCSD1, UCP2, SP140, S1PR1, TLN1, CDK2AP2, CD247, COTL1, LRRFIP1, SLC11A1, SRSF5, CNP, CNN2, KLF6, CLEC4E, INPP5D, GLIPR1, LPCAT2, FERMT3, CLEC4C, FUS, ITGB7, TBC1D10C, NAPSA, PLD4, ADD3, JAK1, AC002996.1, ALDH2, CDKN1B, GSR, MYO1F, CCR7, MXD1, ST8SIA4, GIMAP1-GIMAP5, GIMAP1, GIMAP5, ARPC5, LY6D, RASSF5, IFITM1, IFITM2, IFITM3, ARHGEF1, GDPD3, SMAP2, FAM107B, MCEMP1, BIN2, HDC, FAM32A, POU2F2, KDM6B, CDKN2D, VIM, PAG1, RNF130, SMPDL3A, AC036214.3, TPD52, TMCC1, GRINA, FGL2, CSK, TRBC1, TRBC2, CD27, MACIR, RASAL3, GABARAP, CD28, IPCEF1, RGS19, BCL2, KIAA0040, SLC25A5, RAPGEF6, AC008695.1, PPT1, GNGT2, IRF2, PSME1, CLEC4D, SLC15A3, ANKRD11, PELI1, TSC22D3, RSRP1, YBX1, BANK1, IGLL1, IL7R, CTSA, STAT1, IGSF6, TAX1BP1, NEDD9, TM6SF1, LY86, TRPG1L, HLA- DMA, HLA-DMB, SRRM2, STK4, ABCG1, KLHL6, ABLIM1, RAP1A, EVI2A, BNIP3L, ARHGAP35, LRRC25, OGFRL1, EMP3, CLK1, MKRN1, GCNT2, HACD4, PICALM,WSGR Docket No. 62193-701.601 UBE2D3, ADIPOR1, MAN2B1, CD48, LYL1, PAK2, SEPHS2, LBR, CD69, PHF20L1, SCP2, MS4A1, CDC42SE1, RGS10, STAP1, DDX6, PPP1R2, PPP1R2B, ABI3, ACAP1, PKM, CSF2RB, TNFRSF18, ALOX12, EBF1, LUC7L2, FLNA, IKZF3, PRDX6, SEPTIN1, ARID4A, PI16, GIMAP6, DAZAP2, PPP1R16B, LYST, GRK2, NLRP3, HNRNPA2B1, PTPRCAP, PYCARD, THEMIS2, LBH, MYCBP2, REL, APOBEC1, CD2, CHD2, TREM1, DENND1C, ENTREP3, NCF1, TNFAIP2, PDE4B, ATP6V1G1, CCNL1, FCGR3A, FCGR3B, SKAP2, ELF1, IL2RG, SLC7A11, SMCHD1, CD9, RASGRP1, PBXIP1, PTPN22, MRGPRX3, MRGPRX4, MRGPRX1, MRGPRX2, CMIP, DGKA, IGKC, SLC2A6, ATG3, ARL4C, IL16, BTK, IL21R, PPP1CA, FLI1, GDA, MBD2, MYO1G, PYGL, SCAND1, PTPRE, KDM7A, CTSE, PLBD1, MAP3K3, AKNA, MMP8, YWHAZ, MACROH2A1, SVIL, H2AJ, MFSD6, ZYX, EMILIN2, HNRNPA0, PSENEN, PDCD4, CD5, CHI3L1, PNPLA2, MACF1, NFATC1, PLCG2, RHOF, CALM2, ROCK1, MAP1LC3B2, MAP1LC3B, CAPZB, UBE2B, CCPG1, IL36G, EIF1, GIMAP1-GIMAP5, GIMAP1, GIMAP5, MORF4L1, RSBN1L, CBL, CD300LB, SIGLEC9, SIGLEC7, SIGLEC8, SIGLEC12, SLCO3A1, SUPT4H1, MBP, PPP1R15A, CD79A, NRROS, PPP1R18, PTBP3, GIMAP7, BIRC2, CCDC12, MAPK14, TCF7, C1orf162, RBM25, XRN2, JAKMIP1, DOCK10, VSIR, SIGLEC11, SIGLEC10, RAMP1, CD6, PHIP, TPR, KPNA4, PTAFR, SH3BP5, ITK, TNRC6C, NOTCH2, MYD88, NIPBL, PRKCD, BCL10, IL10RA, DHRS7, CLEC4A, S1PR4, CD300LF, OGT, IRAG2, TTC7A, TREML1, FAU, BCL2A1, MARCKS, MIER1, VASP, CLEC4C, GPCPD1, TRAF3IP3, RUNX3, R3HDM4, LFNG, SLAMF6, PDCD10, STX11, SRSF11, GP5, HNRNPL, C1orf54, PID1, NFE2, MZB1, LILRA5, SCNA, PFDN5, PARP14, VGLL4, ENO1, ARL6IP1, SOCS3, SH2D1A, DAPL1, GPR183, LMNB1, STRBP, IL18RAP, HIPK1, STAG2, ORAI1, SMC6, STK10, FAM111A, GNB2, TREML4, EBI3, AP1S2, ARPC1B, NEURL3, TRIB2, ZC3HAV1, BAZ1A, HECTD1, SEC11C, TNRC6B, PRKD2, RNASE6, EVL, VAV3, RAB27A, TRIM5, ADAM8, PHF3, SIRT7, TCP11L2, ZCCHC7, ATP2B1, ITPR2, BPTF, ACE, ICAM2, SLFN12, SLFN12L, FCMR, PRKAR1A, ASAP1, CLEC2D, MEF2C, KANSL1, SLC40A1, VPS37B, SDCBP, CD8B, CD8B2, PHF11, ZEB2, SFPQ, CCL18, CCL3, CCL3L1, ANXA1, YWHAH, HMGB1, PIK3CD, SERP1, ARRB2, CCDC88B, CTSC, ATF7IP, TREML2, NFKB1, VAMP8, SLC16A3, KIF2A, LMO2, CAPZA2, TBC1D1, GIMAP8, RASSF3, IFNGR2, TONSL, CLEC2D, PTEN, CD226, HLA-DQB1, HLA-DQB2, SGK1, LCN2, RGS14, PTP4A2, TMEM156, MCTP1, ARF1, N4BP1, ARHGAP9, BIRC3, TXK, NPTN, PSME2, ARL6IP5, PSTPIP1, TUBA4A, SERTAD1, DIAPH1, TNFRSF13B, BATF, IL10RB, ARL5C, CCDC88C, CDK9, SH3BP1, GIMAP4, RNF144A, BCL11A, UBE2D2, RASSF2, ARPC3, C19orf53, GRAMD1A, NADK, TNFRSF13C, TRA2A, GGNBP2, ISG15,WSGR Docket No. 62193-701.601 DEF6, STAT4, ATP6V0E1, PRRC2C, CARD19, CLEC12A, GSAP, MFNG, CMTM6, GPR65, DYNLT2B, CNTRL, S100A11, TACC1, TMEM59, ZNF292, SH3KBP1, GPR18, SYF2, POLR1F, RINL, GP9, AIM2, CTCF, CD300E, DCK, OTULIN, PIP4K2A, MAX, TNFSF14, CALM1, KRIT1, MYH9, TMPO, TNF, UBL3, CSNK1A1, CEACAM7, CEACEM5, CEACAM6, CEACAM3, CEACAM1, CEACAM8, BMX, YY1, VRK1, PRKCB, PFKFB4, G3BP2, ABTB1, EPSTI1, RIN3, ITPKB, MYL6, MS4A6A, MS4A6E, LY9, TAP2, PRKAR2B, SF3B6, FAM78A, ABHD8, ZDHHC18, ATP1A3, PPP1R10, MCMBP, TWF2, KRAS, ARHGAP4, TMEM71, ALCAM, NOTCH1, MORC3, SUN2, ALDOA, HAUS8, RSRC2, CDC42EP3, IKBKB, CHD7, ARL11, UBB, CLEC6A, CXCR4, STK24, NFKBIZ, SERINC3, TIMM10B, CTSW, RCHY1, TAP1, MAF1, RBM5, SEMA4A, MAT2B, CYTH1, NFKBID, CD4, GPI, MEF2A, MARCHF2, JUN, GRAP2, AVL9, FCER2, HBA1, HBA2, BCL2A1, ARF6, RNF13, BLNK, IL27RA, KCNN4, RAC1, FPR1, SASH3, FIS1, ATP8B4, SFXN5, GPR141, RSU1, PRR5L, SCNN1A, TMEM243, FCRLA, PIK3R1, NLRP12, MILR1, and any combination thereof.

[0141] In some embodiments, the biological agent recognizes a Coro1a biomarker or a human orthologue of a Coro1a biomarker. In some embodiments, the biological agent recognizes a Cd52 biomarker or a human orthologue of a Cd52 biomarker. In some embodiments, the biological agent recognizes a Cyba biomarker or a human orthologue of a Cyba biomarker. In some embodiments, the biological agent recognizes a Rac2 biomarker or a human orthologue of a Rac2 biomarker. In some embodiments, the biological agent recognizes a Laptm5 biomarker or a human orthologue of a Laptm5 biomarker. In some embodiments, the biological agent recognizes a Ptprc biomarker or a human orthologue of a Ptprc biomarker.

[0142] In some embodiments, one or more maternal biomarkers are bispecific. In some embodiments, the bispecific one or more maternal biomarkers are located in maternal cells and fetal cells. In some embodiments, the bispecific one or more maternal biomarkers are located on maternal cells and fetal cells.

[0143] In some embodiments, the biological agent is a protein. In some embodiments, the biological agent is a ligand. In some embodiments, the biological agent is an antibody. In some embodiments, the antibody has two heavy chains and two light chains. In some embodiments, the heavy chain has a constant region and a variable region. In some embodiments, the light chain has a constant region and a variable region. In some embodiments, the antibody has one heavy chain and one light chain. In some embodiments, the biological agent is an antigen binding fragment of an antibody (Fab). In some embodiments, the Fab has a light chain variable region and a heavy chain variable region. In some embodiments, the Fab has a light chainWSGR Docket No. 62193-701.601 variable region. In some embodiments, the Fab has a light chain variable region and a light chain constant region. In some embodiments, the Fab has a heavy chain variable region. In some embodiments, the Fab has a heavy chain variable region and the heavy chain constant region. In some embodiments, the biological agent is a single chain variable fragment (scFv). In some embodiments, the scFv has a heavy chain variable region and a light chain constant region. In some embodiments, the scFv has a light chain variable region and a heavy chain constant region. In some embodiments, the biological agent is a nanobody. In some embodiments, the nanobody has a light chain variable region. In some embodiments, the nanobody has a heavy chain variable region.

[0144] In some embodiments, the biological agent is configured to couple to a biomarker. In some embodiments, the biological agent is configured to couple directly to a biomarker. In some embodiments, the biological agent couples with a biomarker via a covalent bond. In some embodiments, the covalent bond is a peptide bond. In some embodiments, the biological agent couples to a biomarker via a non-covalent interaction. In some embodiments, the non-covalent interaction is an ionic interaction. In some embodiments, the non-covalent interaction is a hydrogen bond. In some embodiments, the non-covalent interaction is van der Waals interactions. In some embodiments, the non-covalent interaction is a hydrophobic interaction.

[0145] In some embodiments, the biological agent is configured to couple indirectly to a biomarker. In some embodiments, the biological agent couples to the biomarker via a linker. In some embodiments, the linker is a cleavable linker. In some embodiments, the cleavable linker is a chemical labile linker. Non-limiting examples of chemical labile linkers include 6- (maleimidocaproyl)hydrazone, (4-(4-acetylphenoxy)butanoic acid) hydrazone, N-acetyl-γ- calicheamicin dimethyl hydrazide, and any combination thereof. In some embodiments, the cleavable linker is a reducible linker. In some embodiments, the reducible linker is a disulfide linker or a glutathione-sensitive disulfide linker. In some embodiments, the cleavable linker is an enzyme-cleavable linker. Non-limiting examples of enzyme-cleavable linkers include peptide- based linkers, valine-citrulline, phenylalanine-lysine, valine-alanine, para-aminobenzyl carbamate, PEG linkers, β-glucuronide linker, β-galactoside linkers, and any combination thereof. In some embodiments, the cleavable linker is a phosphatase-cleavable linker. In some embodiments, the phosphatase-cleavable linker is a phosphate diester. In some embodiments, the linker is a non-cleavable linker. Non-limiting examples of non-cleavable linkers include thioethers, maleimidocapropy, N-succinimidyl-4-(maleimidomethyl) cyclohexane-1-carboxylate linker, and any combination thereof.WSGR Docket No. 62193-701.601

[0146] In some embodiments, the biological agent is a nucleic acid probe. In some embodiments, the nucleic acid probe is RNA. In some embodiments, the nucleic acid probe is DNA. The nucleic acid probes can be nucleic acid molecules having sequence complementarity with nucleic acid sequences (e.g., RNA or DNA) of the one or more genomic loci (e.g., fetal- associated genomic loci). In some embodiments, the nucleic acid probe can couple to the nucleic acid sequence with sequence complementarity to the nucleic acid probe. In some embodiments, the nucleic acid probe can hybridize to the nucleic acid sequence with sequence complementarity to the nucleic acid probe. These nucleic acid molecules can be primers or enrichment sequences. The assaying of the biological sample using probes that are selective fo r the one or more genomic loci (e.g., fetal-associated genomic loci) can comprise use of array hybridization (e.g., microarray-based), polymerase chain reaction (PCR), or nucleic acid sequencing (e.g., RNA sequencing or DNA sequencing). In some embodiments, DNA or RNA can be assayed by one or more of: isothermal DNA / RNA amplification methods (e.g., loop - mediated isothermal amplification (LAMP), helicase dependent amplification IA), rolling circle amplification (RCA), recombinase polymerase amplification (RPA)), immunoassays, electrochemical assays, surface-enhanced Raman spectroscopy (SERS), quantum dot (QD)- based assays, molecular inversion probes, droplet digital PCR (ddPCR), CRISPR / Cas-based detection (e.g., CRISPR-typing PCR (ctPCR), specific high-sensitivity enzymatic reporter un- locking (SHERLOCK), DNA endonuclease targeted CRISPR trans reporter (DETECTR), and CRISPR-mediated analog multi-event recording apparatus (CAMERA)), and laser transmission spectroscopy (LTS). In some embodiments, the nucleic acid probe can be an aptamer.

[0147] In some embodiments, the biological agent is labeled. In some embodiments, the biological agent is labeled with a protein or peptide. In some embodiments, the peptide is a fluorescent protein or peptide. Non-limiting examples of fluorescent protein or peptide include: CFP (cyan fluorescent protein), YFP (yellow fluorescent protein), mVenus, cyan ECFP (enhanced cyan fluorescent protein), yellow EYFP (enhanced yellow fluorescent protein), green EGFP (enhanced green fluorescent protein), GFP, BPF (blue fluorescent protein), blue EBFP (enhanced blue fluorescent protein), red mCherry, SYTO®9, propidium iodide, mCitrine, YPet, aquamarine, mTurquoise2, mCeruean3, LUMP (lumazine binding protein), mTFP1 (monomeric teal fluorescent protein), NowGFP, Clover, mClover3, mNeonGreen, mRuby2 mRuby3, mPlum, eqFP650, mCardinal, IFP1.4m, iRFPm, mAmetrine, LSS-mOrange, tdTomato, mKate2, ShadowG, REACh1 (Resonance Energy-Accepting Chromoprotein 1), REACh2, sREACh, rsTagRFP, PA-GFP (photo activatable green fluorescent protein), Phanta, T-Sapphire, mTagBFP, sfGFP (superfolder GFP), CyOFP1, mOrange2, mKOκ, TagRFP, DsRed, muGFP,WSGR Docket No. 62193-701.601 AF660 (Alex Fluor 660),hydrxycoumarin, aminocoumarin, methoxycoumarin, cascade blue, lucifer yellow, NBD, R-Phycoerythrin, PE-Cy5, PE-Cy7, APC-Cy7, fluorescein, FluorX, BODIPY-FL, tetramethylrhodamine, tetramethylrhodamine isothiocyanate, X-Rhodamine, Lissamine, Rhodamine B, PerCP, Texas Red, Allophycocyanin, TruRed, Alexa Fluor 350, Alexa Fluor 430, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluo r 555, Alexa Fluor 568, Alexa Fluor 594, Alexa Flour 633, Alexa Fluor 647, Alexa Fluor 660, Alexa Flour 680, Alexa Fluor 700, Alexa Fluor 750, SpectrumOrange, SpectrumGreen1, SpectrumGreen2, SpectrumAqua, SpectrumBlue, SpectrumGold, SpectrumRed, SpectrumFRed, Cy2, Cy3, Cy3.5, Cy5, Cy5.5, Cy7, or Ni-NTA-Atto, and fragments thereof.

[0148] In some embodiments, the biological agent is coupled with a fluorescent protein or peptide. In some embodiments, the biological agent is coupled with a fluorescent protein or peptide via a covalent bond. In some embodiments, the covalent bond is a peptide bond. In some embodiments, the biological agent is coupled to a fluorescent protein or peptide via a non - covalent interaction. In some embodiments, the non-covalent interaction is an ionic bond. In some embodiments, the non-covalent interaction is a hydrogen bond. In some embodiments, the non-covalent interaction is van der Waals interactions. In some embodiments, the non-covalent interaction is a hydrophobic interaction.

[0149] In some embodiments, the biological agent is indirectly coupled with a fluorescent protein or peptide. In some embodiments, the biological agent is indirectly coupled with a fluorescent protein or peptide via a linker. In some embodiments, the linker is a short amino acid sequence. In some embodiments, the short amino acid sequence is from one amino acid residue to about 10 amino acid residues. In some embodiments, the short amino acid sequence is made up of glycine residues. In some embodiments, the short amino acid sequence is made up of serine residues. In some embodiments, the short amino acid sequence is made up of glycine and serine residues. In some embodiments, the linker is polyethylene glycol. In some embodiments, the linker is streptavidin. In some embodiments, the linker is an alkylene group. The linker can be flexible, rigid, and / or cleavable.

[0150] The linker can comprise a polymeric spacer, a polyether spacer, and / or an alkylene spacer (e.g., methylene, ethylene, propylene, or butylene). In some embodiments, the linker can be a bond (e.g., a covalent bond). The spacer can comprise polyethylene glycol (PEG) groups, alkylene groups, and / or amino acid residues.

[0151] In some embodiments, the linker can have from about 1 PEG group to about 10 PEG groups. In some embodiments, the linker can have at least about 1 PEG group, at least about 2 PEG groups, at least about 3 PEG groups, at least about 4 PEG groups, at least about 5 PEGWSGR Docket No. 62193-701.601 groups, at least about 6 PEG groups, at least about 7 PEG groups, at least about 8 PEG groups, at least about 9 PEG groups, at least about 10 PEG groups, or more. In some embodiments, the linker can have at most about 10 PEG groups, at most about 9 PEG groups, at most about 8 PEG groups, at most about 7 PEG groups, at most about 6 PEG groups, at most about 5 PEG groups, at most about 4 PEG groups, at most about 3 PEG groups, at most about 2 PEG groups, or at most about 1 PEG groups. In some embodiments, the linker can have about 1 PEG group, about 2 PEG groups, about 3 PEG groups, about 4 PEG groups, about 5 PEG groups, about 6 PEG groups, about 7 PEG groups, about 8 PEG groups, about 9 PEG groups, or about 10 PEG groups.

[0152] In some embodiments, the linker can have from about 1 to about 10 alkylene groups. In some embodiments, the linker can have at least about 1 alkylene group, at least about 2 alkylene groups, at least about 3 alkylene groups, at least about 4 alkylene groups, at least about 5 alkylene groups, at least about 6 alkylene groups, at least about 7 alkylene groups, at least about 8 alkylene groups, at least about 9 alkylene groups, at least about 10 alkylene groups, or more. In some embodiments, the linker can have at most about 10 alkylene groups, at most about 9 alkylene groups, at most about 8 alkylene groups, at most about 7 alkylene groups, at most about 6 alkylene groups, at most about 5 alkylene groups, at most about 4 alkylene groups, at most about 3 alkylene groups, at most about 2 alkylene groups, at most about 1 alkylene group, or less. In some embodiments, the linker can have about 1 alkylene group, about 2 alkylene groups, about 3 alkylene groups, about 4 alkylene groups, about 5 alkylene groups, about 6 alkylene groups, about 7 alkylene groups, about 8 alkylene groups, about 9 alkylene groups, or about 10 alkylene groups.

[0153] In some embodiments, the linker can have from about 1 amino acid residue to about 10 amino acid residues. In some embodiments, the linker can have at least about one amino acid residue, at least about two amino acid residues, at least about three amino acid residues, at least about four amino acid residues, at least about five amino acid residues, at least about six amino acid residues, at least about seven amino acid residues, at least about eight amino acid residues, at least about nine amino acid residues, at least about ten amino acid residues, or more. In some embodiments, the linker can have at most about ten amino acid residues, at most about nine amino acid residues, at most about eight amino acid residues, at most about seven amino acid residues, at most about six amino acid residues, at most about five amino acid residues, at most about four amino acid residues, at most about three amino acid residues, at most about two amino acid residues, or at most about one amino acid residue. In some embodiments, the linker can have about one amino acid residue, about two amino acid residues, about three amino acidWSGR Docket No. 62193-701.601 residues, about four amino acid residues, about five amino acid residues, about six amino acid residues, about seven amino acid residues, about eight amino acid residues, about nine amino acid residues, or about ten amino acid residues.

[0154] In some embodiments, the biological agent is attached to a surface. In some embodiments, the surface is a plate. In some embodiments, the surface is a slide. In some embodiments, the surface is a bead. In some embodiments, the bead is plastic. In some embodiments, the bead is glass. In some embodiments, the bead is metal. In some embodiments, the bead is magnetic.

[0155] In some embodiments, one or more biological agents are attached to the surface. In some embodiments, up to about 10 biological agents are attached to the surface. In some embodiments, at least about one biological agents, at least about two biological agents, at least about three biological agents, at least about four biological agents, at least about five biological agents, at least about six biological agents, at least about seven biological agents, at least about eight biological agents, at least about nine biological agents, or at least about ten biological agents are attached to the surface.

[0156] In some embodiments, at most about ten biological agents, at most about nine biological agents, at most about eight biological agents, at most about seven biological agents, at most about six biological agents, at most about five biological agents, at most about four biological agents, at most about three biological agents, at most about two biological agents, or at most about one biological agent is attached to the surface.

[0157] In some embodiments, about one biological agent, about two biological agents, about three biological agents, about four biological agents, about five biological agents, about six biological agents, about seven biological agents, about eight biological agents, about nine biological agents, or about ten biological agents are attached to the surface.

[0158] In some embodiments, the one or more biological agents (e.g., isolation agents or agents) that specifically couple (e.g., are specific for) to one or more biomarkers couple to the one or more biomarkers. In some embodiments, the one or more biological agents that are specific for the one or more biomarkers do not couple to other biomarkers that are different from the one or more biomarkers.

[0159] In some embodiments, the one or more biological agents (e.g., isolation agents) are biological agents that specifically couple to one or more fetal biomarkers. In some embodiments, the one or more biological agents (e.g., isolation agents) that specifically couple (e.g., are specific for) to one or more fetal biomarkers couple to the one or more fetal biomarkers. In some embodiments, the one or more biological agents that are specific for the one or more fetalWSGR Docket No. 62193-701.601 biomarkers do not couple to other fetal biomarkers that are different from the one or more fetal biomarkers. In some embodiments, the one or more biological agents that are specific for the one or more fetal biomarkers may not couple to one or more maternal biomarkers.

[0160] In some embodiments, the one or more biological agents are biological agents that specifically couple to one or more maternal biomarkers. In some embodiments, the one or more biological agents (e.g., isolation agents) that specifically couple (e.g., are specific for) to one or more maternal biomarkers couple to the one or more maternal biomarkers. In some embodiments, the one or more biological agents that are specific for the one or more maternal biomarkers may not couple to other maternal biomarkers that are different from the one or more maternal biomarkers. In some embodiments, the one or more biological agents that are specific for the one or more maternal biomarkers may not couple to one or more fetal biomarkers. Methods

[0161] In some embodiments, the present disclosure provides a method of analyzing a nucleic acid sequence. In some embodiments, the nucleic acid sequence is DNA. In some embodiments, the DNA is genomic DNA. In some embodiments, the nucleic acid sequence is RNA. In some embodiments, the RNA is mRNA. In some embodiments, the RNA is rRNA. In some embodiments, the RNA is lncRNA. In some embodiments, the nucleic acid sequence is isolated from a biological sample. In some embodiments, the biological sample is a blood sample. In some embodiments, the blood sample is blood plasma. In some embodiments, the blood sample is blood serum. In some embodiments, the blood sample is a buffy coat. In some embodiments, the blood sample is peripheral blood mononuclear cells. In some embodiments, the blood sample is nucleated white blood cells. In some embodiments, the blood sample is whole blood. In some embodiments, the blood sample is whole blood depleted of red blood cell. In some embodiments, the biological sample is a tissue and / or biopsy sample. In some embodiments, the tissue and / or biopsy sample is from the uterus, placenta, umbilical cord, or any combination thereof. In some embodiments, the biological sample is a biological fluid sample. In some cases, the biological fluid sample is blood, saliva, interstitial fluid, umbilical cord blood, extracellular fluid, or any combination thereof. In some embodiments, the sample is from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a mouse. In some embodiments, the subject is a rat. In some embodiments, the subject is a non-human primate. In some embodiments, the subject is a human. In some embodiments, the sample is from a pregnant subject. In some embodiments, the sample is a maternal biological sample. In some embodiments, the biological blood sample is from a pregnant subject.WSGR Docket No. 62193-701.601

[0162] In some embodiments, the nucleic acid sequence is from an ex vivo population of cells. In some embodiments, the ex vivo population of cells has a subpopulation of fetal cells. In some embodiments, the ex vivo population of cells has a subpopulation of maternal cells. In some embodiments, the ex vivo population of cells has a subpopulation of fetal cells and a subpopulation of maternal cells.

[0163] In some embodiments, the method further comprising depleting red blood cells from the ex vivo population of cells. In some embodiments, red blood cells are depleted from the ex vivo population of cells to remove non-nucleated cells from the sample. In some embodiments, red blood cells are depleted from the ex vivo population of cells to decrease background interference in subsequent analyses and / or assays performed on the ex vivo population of cells. In some embodiments, red blood cells are depleted from the ex vivo population of cells to increase the accuracy in subsequent analyses and / or assays performed on the ex vivo population of cells. In some embodiments, red blood cells (RBCs) are depleted from the ex vivo population of cells via ammonium chloride lysis, density gradient centrifugation, hypotonic lysis, immunomagnetic cell separation, sedimentation, or any combination thereof.

[0164] In some embodiments, the method further comprises providing an extract prior to sequencing. In some embodiments, the extract is made by extracting the ex vivo population of cells from the maternal biological sample. In some embodiments, the ex vivo population of cells is extracted from the maternal biological sample. In some embodiments, the subpopulation of fetal cells is extracted from the maternal biological sample. In some embodiments, the subpopulation of maternal cells is extracted from the maternal biological sample. In some embodiments, the subpopulation of maternal cells and the subpopulation of fetal cells are extracted from the maternal biological sample.

[0165] In some embodiments, the subpopulation of fetal cells has from about one fetal cell type to about ten fetal cell types. Non-limiting examples of fetal cell types include early gestation trophoblasts, late gestation trophoblasts, erythroblasts, megakaryocytes, stromal cells, endothelial cells, lymphocytes, nucleated red blood cells, hematopoietic progenitor cells, vascular endothelial cells, and mesenchymal progenitor cells. In some embodiments, the subpopulation of fetal cells has at least about one fetal cell type, at least about two fetal cell types, at least about three fetal cell types, at least about four fetal cell types, at least about five fetal cell types, at least about six fetal cell types, at least seven fetal cell types, at least about eight fetal cell types, at least about nine fetal cell types, or more. In some embodiments, the subpopulation of fetal cells has at most about ten fetal cell types, at most about nine fetal cell types, at most about eight fetal cell types, at most about seven fetal cell types, at most about sixWSGR Docket No. 62193-701.601 fetal cell types, at most about five fetal cell types, at most about four fetal cell types, at most about three fetal cell types, at most about two fetal cell types, at most about one fetal cell type, or less. In some embodiments, the subpopulation of fetal cells has about one fetal cell type, about two fetal cell types, about three fetal cell types, about four fetal cell types, about five fetal cell types, about six fetal cell types, about seven f etal cell types, about eight fetal cell types, about nine fetal cell types, or about ten fetal cell types.

[0166] In some embodiments, the subpopulation of maternal cells has from about one maternal cell type to about 10 maternal cell types. In some embodiments, the subpopulation of maternal cells has at least about one maternal cell types, at least about two maternal cell types, at least about three maternal cell types, at least about four maternal cell types, at least about five maternal cell types, at least about six maternal cell types, at least about seven maternal cell types, at least about eight maternal cell types, at least about nine maternal cell types, at least about ten maternal cell types, or more. In some embodiments, the subpopulation of maternal cells has at most about ten maternal cell types, at most about nine maternal cell types, at most about eight maternal cell types, at most about seven maternal cell types, at most about six maternal cell types, at most about five maternal cell types, at most about four maternal cell types, at most about three maternal cell types, at most about two maternal cell types, at most about one maternal cell type, or less. In some embodiments, the subpopulation of maternal cells has about one maternal cell types, about two maternal cell types, about three maternal cell types, about four maternal cell types, about five maternal cell types, about six maternal cell types, about seven maternal cell types, about eight maternal cell types, about nine maternal cell types, or about ten maternal cell types.

[0167] In some embodiments, the ex vivo population of cells is extracted from a mother. In some embodiments, the ex vivo population of cells is extracted from a mother by taking a biological sample. In some embodiments, the ex vivo population of cells is extracted from a mother by taking a biological fluid sample. In some embodiments, the ex vivo population of cells is extracted from a mother by taking a blood sample. In some embodiments, the ex vivo population of cells is extracted from a mother by taking a tissue sample and / or biopsy. In some embodiments, the ex vivo population of cells has an ex vivo population of fetal cells. In some embodiments, the ex vivo population of cells has an ex vivo population of maternal cells. In some embodiments, the ex vivo population of cells has an ex vivo population of the fetal cells and an ex vivo population of maternal cells.

[0168] In some embodiments, the ex vivo population of cells is extracted from the biological sample. In some cases, the ex vivo population of cells is extracted from the biological sample byWSGR Docket No. 62193-701.601 removing non-nucleated cells (e.g., red blood cells), extracellular proteins, extracellular nucleic acid molecules, extracellular metabolites, or any combination thereof from the biological sample.

[0169] In some embodiments, subsequent to extracting the ex vivo population of cells from the biological sample, the ex vivo population of cells can undergo one or more washing operations. In some embodiments, the one or more washing operations can eliminate extracellular molecules (e.g., lipids, sugars, minerals, vitamins, nucleic acids, metabolites) remaining on the cells. In some embodiments, the one or more washing operations can remove excess reagents and / or labels. In some embodiments, the method can comprise from about one to about two wash operations, from about two to about three wash operations, from about three to about four wash operations, from about four wash operations to about five wash operations, from about five wash operations to about six wash operations, from about six wash operations to about seven wash operations, from about seven wash operations to about eight wash operations, from about eight wash operations to about nine wash operations, or from about nine wash operations to about ten wash operations. In some embodiments, the method can comprise at least about one wash operation, at most about two wash operations, at least about three wash operations, at least about four wash operations, at least about five wash operations, at least about six wash operations, at least about seven wash operations, at least about eight wash operations, at least about nine wash operations, at least about ten wash operations or more. In some cases, the method can comprise at most about ten wash operations, at most about nine wash operations, at most about eight wash operations, at most about seven wash operations, at most about six wash operations, at most about five wash operations, at most about four wash operations, at most about three wash operations, at most about two wash operations, at most about one wash operation, or less. In some cases, the method can comprise about one wash operation, about two wash operations, about three wash operations, about four wash operations, about five wash operations, about six wash operations, about seven wash operations, about eight wash operations, about nine wash operations, or about ten wash operations. In some embodiments, the washing operation uses a washing buffer. Non-limiting examples of washing buffers include water, methanol, ethanol, phosphate buffered saline (PBS), saline, Tris-buffered solution, Tween buffer, glycine buffer, and any combination thereof.

[0170] In some embodiments, the ex vivo population of cells extracted from the maternal biological sample is used to provide an extract. In some embodiments, the ex vivo population of cells is extracted from the maternal biological sample using density gradient centrifugation. In some embodiments, the ex vivo population of cells is extracted from the maternal biologicalWSGR Docket No. 62193-701.601 sample using magnetic isolation. In some embodiments, the ex vivo population of cells is extracted from the maternal biological sample using shear-induced diffusion. In some embodiments, the extract is made up of the subpopulation of fetal cells. In some embodiments, the extract is made up of the subpopulation of maternal cells. In some embodiments, the extract is made up of the subpopulation of fetal cells and the subpopulation of maternal cells.

[0171] In some embodiments, the method further comprises enriching the subpopulation of fetal cells in the extract to provide an enriched sample. In some embodiments, the extract is enriched to provide an enriched sample. In some embodiments, the enriched sample is enriched in fetal cells. In some embodiments, the enriched sample is depleted in maternal cells. In some embodiments, the enriched sample is enriched in fetal cells and depleted in maternal cells. In some embodiments, the enriched sample is depleted in maternal cells and subsequently enriched for fetal cells. In some embodiments, the enriched sample is enriched for fetal cells and subsequently depleted for maternal cells. In some embodiments, the extract is enriched for fetal cells by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the extract is enriched for fetal cells by fluorescence-activated cell sorting (FACS). In some embodiments, the extract is enriched for fetal cells by magnetic-activated cell sorting (MACS). In some embodiments, the extract is enriched for fetal cells using one or more fetal cell biomarkers. In some embodiments, the extract is enriched for fetal cells by negative immunomagnetic cell separation. In some embodiments, the negative immunomagnetic cell separation enriches for fetal cells using one or more maternal cell biomarkers. In some embodiments, the extract is enriched for fetal cells by microscopic microdissection. In some embodiments, the extract is enriched for fetal cells by microscopic imaging.

[0172] In some embodiments, the extract is enriched for fetal cells using one or more isolation agents. In some embodiments, the extract is enriched for fetal cells by incubating one or more isolation agents with the ex vivo population of cells in the extract. In some embodiments, the one or more isolation agents incubated with the ex vivo population of cells in the extract are specific for one or more fetal biomarkers. In some embodiments, the one or more isolation agents incubated with the ex vivo population of cells in the extract are specific for one or more maternal biomarkers. In some embodiments, the one or more isolation agents incubated with the ex vivo population of cells in the extract are specific for one or more fetal biomarkers and specific for one or more maternal biomarkers.

[0173] In some embodiments, subsequent to incubating (e.g., contacting) the ex vivo population of cells with the one or more isolation agents, the ex vivo population of fetal cells can undergoWSGR Docket No. 62193-701.601 one or more washing operations. In some embodiments, the one or more washing operations can eliminate any unbound isolation agents. In some embodiments, the one or more washing operations can remove excess reagents and / or labels. In some embodiments, the method can comprise from about one to about two wash operations, from about two to about three wash operations, from about three to about four wash operations, from about four wash operations to about five wash operations, from about five wash operations to about six wash operations, from about six wash operations to about seven wash operations, from about seven wash operations to about eight wash operations, from about eight wash operations to about nine wash operations, or from about nine wash operations to about ten wash operations. In some embodiments, the method can comprise at least about one wash operation, at most about two wash operations, at least about three wash operations, at least about four wash operations, at least about five wash operations, at least about six wash operations, at least about seven wash operations, at least about eight wash operations, at least about nine wash operations, at least about ten wash operations or more. In some embodiments, the method can comprise at most about ten wash operations, at most about nine wash operations, at most about eight wash operations, at most about seven wash operations, at most about six wash operations, at most about five wash operations, at most about four wash operations, at most about three wash operations, at most about two wash operations, at most about one wash operation, or fewer. In some embodiments, the method can comprise about one wash operation, about two wash operations, about three wash operations, about four wash operations, about five wash operations, about six wash operations, about seven wash operations, about eight wash operations, about nine wash operations, or about ten wash operations.

[0174] In some embodiments, the extract is sorted to isolate the ex vivo population of fetal cells. In some embodiments, the extract is sorted to separate the ex vivo population of fetal cells from the ex vivo population of maternal cells from the extract. In some embodiments, the extract is sorted for fetal cells by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the extract is sorted for fetal cells by fluorescence-activated cell sorting (FACS). In some embodiments, the extract is sorted for fetal cells by magnetic-activated cell sorting (MACS). In some embodiments, the extract is sorted for fetal cells using a presence of one or more fetal cell biomarkers. In some embodiments, the extract is sorted for fetal cells by negative immunomagnetic cell separation. In some embodiments, the negative immunomagnetic cell separation sorts fetal cells using an absence of one or more maternal cell biomarkers. In someWSGR Docket No. 62193-701.601 embodiments, the extract is sorted for fetal cells by microscopic microdissection. In some embodiments, the extract is sorted for fetal cells by microscopic imaging.

[0175] In some embodiments, the extract is sorted to isolate the ex vivo population of maternal cells. In some embodiments, the extract is sorted to separate the ex vivo population of fetal cells from the ex vivo population of maternal cells from the extract. In some embodiments, the extract is sorted for maternal cells by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the extract is sorted for maternal cells by fluorescence-activated cell sorting (FACS). In some embodiments, the extract is sorted for maternal cells by magnetic-activated cell sorting (MACS). In some embodiments, the extract is sorted for maternal cells using a presence of one or more fetal cell biomarkers. In some embodiments, the extract is sorted for maternal cells by negative immunomagnetic cell separation. In some embodiments, the negative immunomagnetic cell separation sorts maternal cells using an absence of one or more fetal cell biomarkers. In some embodiments, the extract is sorted for maternal cells by microscopic microdissection. In some embodiments, the extract is sorted for maternal cells by microscopic imaging.

[0176] In some embodiments, the ex vivo population of fetal cells is isolated from the extract. In some embodiments, the ex vivo population of fetal cells is separated from the extract. In some embodiments, the fetal cells are isolated from the extract by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the fetal cells are isolated from the extract by fluorescence- activated cell sorting (FACS). In some embodiments, the fetal cells are isolated from the extract by magnetic-activated cell sorting (MACS). In some embodiments, the fetal cells are isolated from the extract using a presence of one or more fetal cell biomarkers. In some embodiments, the fetal cells are isolated from the extract by negative immunomagnetic cell separation. In some embodiments, the negative immunomagnetic cell separation isolates fetal cells using an absence of one or more maternal cell biomarkers. In some embodiments, the fetal cells are isolated from the extract by microscopic microdissection. In some embodiments, the fetal cells are isolated from the extract by microscopic imaging.

[0177] In some embodiments, the ex vivo population of maternal cells is isolated from the extract. In some embodiments, the ex vivo population of maternal cells is separated from the extract. In some embodiments, the maternal cells are isolated from the extract by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the maternal cells are isolatedWSGR Docket No. 62193-701.601 from the extract by fluorescence-activated cell sorting (FACS). In some embodiments, the maternal cells are isolated from the extract by magnetic-activated cell sorting (MACS). In some embodiments, the maternal cells are isolated from the extract using a presence of one or more maternal cell biomarkers. In some embodiments, the maternal cells are isolated from the extract by negative immunomagnetic cell separation. In some embodiments, the negative immunomagnetic cell separation isolates maternal cells using an absence of one or more fetal cell biomarkers. In some embodiments, the maternal cells are isolated from the extract by microscopic microdissection. In some embodiments, the maternal cells are isolated from the extract by microscopic imaging.

[0178] In some embodiments, the extract is enriched for fetal cells using from about one isolation agent to about five isolation agents, or from about five isolation agents to about ten isolation agents. In some embodiments, the extract is enriched for fetal cells using at least about one isolation agent, at least about two isolation agents, at least about three isolation agents, at least about four isolation agents, at least about five isolation agents, at least about six isolation agents, at least about seven isolation agents, at least about eight isolation agents, at least about nine isolation agents, at least about ten isolation agents, or more. In some embodiments, the extract is enriched for fetal cells using at most about ten isolation agents, at most about nine isolation agents, at most about eight isolation agents, at most about seven isolation agents, at most about six isolation agents, at most about five isolation agents, at most about four isolation agents, at most about three isolation agents, at most about two isolation agents, at most about one isolation agent, or less. In some embodiments, the extract is enriched for fetal cells using about one isolation agent, about two isolation agents, about three isolation agents, about four isolation agents, about five isolation agents, about six isolation agents, about seven isolation agents, about eight isolation agents, about nine isolation agents, or about ten isolation agents.

[0179] In some embodiments, the one or more isolation agents has a first isolation agent. In some embodiments, the first isolation agent couples to a first biomarker in the subpopulation of fetal cells. In some embodiments, the extract is enriched for fetal cells by density gradient centrifugation. In some embodiments, the extract is enriched for fetal cells by immunodensity cell isolation. In some embodiments, the extract is enriched for fetal cells by microfluidic cell sorting.

[0180] In some embodiments, the method further comprises enriching the subpopulation of maternal cells in the extract to provide an enriched sample. In some embodiments, the extract is enriched to provide an enriched sample. In some embodiments, the enriched sample is enriched in maternal cells. In some embodiments, the enriched sample is depleted in fetal cells. In someWSGR Docket No. 62193-701.601 embodiments, the enriched sample is enriched in maternal cells and depleted in fetal cells. In some embodiments, the enriched sample is depleted in fetal cells and subsequently enriched for maternal cells. In some embodiments, the enriched sample is enriched for maternal cells and subsequently depleted in fetal cells. In some embodiments, extract is enriched for maternal cells by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the extract is enriched for maternal cells by fluorescence-activated cell sorting (FACS). In some embodiments, the extract is enriched for maternal cells by magnetic-activated cell sorting (MACS). In some embodiments, the extract is enriched for maternal cells by microscopic microdissection. In some embodiments, the extract is enriched for maternal cells by microscopic imaging. In some embodiments, the extract is enriched for maternal cells using one or more maternal cell biomarkers. In some embodiments, the extract is enriched for maternal cells by negative immunomagnetic cell separation. In some embodiments, the negative immunomagnetic cell separation enriches for maternal cells using one or more fetal cell biomarkers.

[0181] In some embodiments, the extract is enriched for maternal cells using one or more isolation agents. In some embodiments, the extract is enriched for maternal cells using from about one isolation agent to about five isolation agents, or from about five isolation agents to about ten isolation agents. In some embodiments, the extract is enriched for maternal cells using at least about one isolation agent, at least about two isolation agents, at least about three isolation agents, at least about four isolation agents, at least about five isolation agents, at least about six isolation agents, at least about seven isolation agents, at least about eight isolation agents, at least about nine isolation agents, at least about ten isolation agents, or more. In some embodiments, the extract is enriched for maternal cells using at most about ten isolation agents, at most about nine isolation agents, at most about eight isolation agents, at most about seven isolation agents, at most about six isolation agents, at most about five isolation agents, at most about four isolation agents, at most about three isolation agents, at most about two isolation agents, at most about one isolation agent, or less. In some embodiments, the extract is enriched for maternal cells using about one isolation agent, about two isolation agents, about three isolation agents, about four isolation agents, about five isolation agents, about six isolation agents, about seven isolation agents, about eight isolation agents, about nine isolation agents, or about ten isolation agents. In some embodiments, the one or more isolation agents has a second isolation agent. In some embodiments, the extract is enriched for maternal cells using one or more isolation agent. In some embodiments, the one or more isolation agent couples to a second biomarker in the subpopulation of maternal cells.WSGR Docket No. 62193-701.601

[0182] In some embodiments, the method further comprises sorting the subpopulation of the fetal cells in the extract from the subpopulation of maternal cells. In some embodiments, the sorting of the subpopulation of the cells in the extract provides an enriched sample. In some embodiments, the extract is sorted to provide an enriched sample. In some embodiments, the enriched sample is enriched in fetal cells. In some embodiments, the enriched sample is depleted in maternal cells. In some embodiments, the enriched sample is enriched in fetal cells and depleted in maternal cells. In some embodiments, the extract is enriched for fetal cells by sorting the extract for a cell characteristic. Non-limiting examples of cell characteristics include size, expression of a specific gene, expression of an extracellular protein, and expression of an intracellular protein.

[0183] In some embodiments, the extract is enriched for fetal cells by sorting the extract for cells that are from about 6 micrometers (μm) to about 14 μm in diameter. In some embodiments, the extract is enriched for fetal cells by sorting the extract for cells that are at least about 6 μm, at least about 7 μm, at least about 8 μm, at least about 9 μm, at least about 10 μm, at least about 11 μm, at least about 12 μm, at least about 13 μm, at least about 14 μm, or more in diameter. In some embodiments, the extract is enriched for fetal cells by sorting the extract for ce lls that are at most about 14 μm, at most about 13 μm, at most about 12 μm, at most about 11 μm, at most about 10 μm, at most about 9 μm, at most about 8 μm, at most about 7 μm, at most about 6 μm, or less in diameter. In some embodiments, the extract is enriched for fetal cells by sorting the extract for cells that are about 6 μm, about 7 μm, about 8 μm, about 9 μm, about 10 μm, about 11 μm, about 12 μm, about 13 μm, or about 14 μm in diameter.

[0184] In some embodiments, the extract is enriched for fetal cells by sorting the extract for expression of an extracellular biomarker. In some embodiments, the extract is enriched for fetal cells by sorting the extract for the absence of expression of an extracellular biomarker. In some embodiments, the extract is enriched for fetal cell by sorting the extract for expression of an intracellular biomarker. In some embodiments, the extract is enriched for fetal cells by sorting the extract for the absence of expression of an intracellular biomarker. In some embodiments, the extract is enriched for fetal cells by sorting the extract for expression of a specific gene. In some embodiments, the extract is enriched for fetal cells by sorting the extract for absence of expression of a specific gene. In some embodiments, the specif ic gene is expressed as DNA. In some embodiments, the specific gene is expressed as RNA. In some embodiments, the extract is sorted by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the extract is enriched for fetal cells by negative immunomagnetic cell separation. In some embodiments, theWSGR Docket No. 62193-701.601 negative immunomagnetic cell separation enriches for fetal cells using one or more maternal cell biomarkers. In some embodiments, the extract is sorted by FACS. In some embodiments, the extract is sorted by MACS. In some embodiments, the extract is sorted by microscopic microdissection. In some embodiments, the extract is sorted by microscopic imaging. In some embodiments, the extract is sorted for fetal cells using one or more isolation agent. In some embodiments, the one or more isolation agents has a first isolation agent. In some embodiments, the first isolation agent couples to a first biomarker in the subpopulation of fetal cells. In some embodiments, the extract is sorted by microfluidic cell sorting.

[0185] In some embodiments, the method further comprises sorting the subpopulation of the maternal cells in the extract from the subpopulation of fetal cells. In some embodiments, the sorting of the subpopulation of the cells in the extract provides an enriched sample. In some embodiments, the extract is sorted to provide an enriched sample. In some embodiments, the enriched sample is enriched in maternal cells. In some embodiments, the enriched sample is depleted in fetal cells. In some embodiments, the enriched sample is enriched in maternal cells and depleted in fetal cells. In some embodiments, the extract is enriched for maternal cells by sorting the extract for a cell characteristic. Non-limiting examples of cell characteristics include size, expression of a specific gene, expression of an extracellular protein, and expression of an intracellular protein.

[0186] In some embodiments, the extract is enriched for maternal cells by sorting the extract for cells that are from about 6 micrometers (μm) to about 14 μm in diameter. In some embodiments, the extract is enriched for maternal cells by sorting the extract for cells that are at least about 6 μm, at least about 7 μm, at least about 8 μm, at least about 9 μm, at least about 10 μm, at least about 11 μm, at least about 12 μm, at least about 13 μm, at least about 14 μm, or more in diameter. In some embodiments, the extract is enriched for maternal cells by sorting the extract for cells that are at most about 14 μm, at most about 13 μm, at most about 12 μm, at most about 11 μm, at most about 10 μm, at most about 9 μm, at most about 8 μm, at most about 7 μm, at most about 6 μm, or less in diameter. In some embodiments, the extract is enriched for maternal cells by sorting the extract for cells that are about 6 μm, about 7 μm, about 8 μm, about 9 μm, about 10 μm, about 11 μm, about 12 μm, about 13 μm, or about 14 μm in diameter.

[0187] In some embodiments, the extract is enriched for maternal cells by sorting the extract for expression of an extracellular biomarker. In some embodiments, the extract is enriched for maternal cells by sorting the extract for the absence of expression of an extracellular biomarker. In some embodiments, the extract is enriched for maternal cell by sorting the extract for expression of an intracellular biomarker. In some embodiments, the extract is enriched forWSGR Docket No. 62193-701.601 maternal cells by sorting the extract for the absence of expression of an intracellular biomarker. In some embodiments, the extract is enriched for maternal cells by sorting the extract for expression of a specific gene. In some embodiments, the extract is enriched for maternal cells by sorting the extract for absence of expression of a specific gene. In some embodiments, the specific gene is expressed as DNA. In some embodiments, the specific gene is expressed as RNA. In some embodiments, the extract is sorted by immunomagnetic cell separation. In some embodiments, the immunomagnetic cell separation is positive immunomagnetic cell selection. In some embodiments, the extract is enriched for maternal cells by negative immunomagnetic cell separation. In some embodiments, the negative immunomagnetic cell separation enriches for maternal cells using one or more fetal cell biomarkers. In some embodiments, the extract is sorted by FACS. In some embodiments, the extract is sorted by MACS. In some embodiments, the extract is sorted by microscopic microdissection. In some embodiments, the extract is sorted by microscopic imaging. In some embodiments, the extract is sorted for maternal cells using one or more isolation agents. In some embodiments, the one or more isolation agents has a second isolation agent. In some embodiments, the second isolation agent couples to a second biomarker in the subpopulation of maternal cells.

[0188] In some embodiments, the subpopulation of fetal cells is early gestation fetal cells. In some embodiments, the subpopulation of fetal cells is later gestation fetal cells. In some embodiments, the subpopulation of fetal cells is trophoblasts. In some embodiments, the subpopulation of fetal cells is early gestation trophoblasts. In some embodiments, the subpopulation of fetal cells is later gestation trophoblasts. In some embodiments, the subpopulation of fetal cells is erythroblasts. In some embodiments, the subpopulation of fetal cells is megakaryocytes. In some embodiments, the subpopulation of fetal cells is stromal cells. In some embodiments, the subpopulation of fetal cells is endothelial cells. In some embodiments, the subpopulation of fetal cells is lymphocytes. In some embodiments, the subpopulation of fetal cells is nucleated red blood cells. In some embodiments, the subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments , the subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the subpopulation of fetal cells is vascular endothelial cells.

[0189] In some embodiments, the subpopulation of fetal cells is made up for one or more subpopulations of fetal cells. In some embodiments, the second subpopulation of fetal cells is different in cell type from the first subpopulation of fetal cells. In some embodiments, the first subpopulation of fetal cells is early gestation fetal cells and the second subpopulation of fetal cells is later gestation fetal cells. In some embodiments, the first subpopulation of fetal cells isWSGR Docket No. 62193-701.601 early gestation trophoblasts and the second subpopulation of fetal cells is later gestation trophoblasts. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is early gestation trophoblasts.

[0190] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is erythroblasts. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is erythroblasts.

[0191] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is megakaryocytes. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is megakaryocytes.

[0192] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is stromal cells. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopu lation of fetal cells is stromal cells.

[0193] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is endothelial cells. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is endothelial cells.

[0194] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is lymphocytes. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is lymphocytes.

[0195] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is nucleated red blood cells.

[0196] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is hematopoietic progenitor cells.

[0197] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is mesenchymal progenitor cells. In someWSGR Docket No. 62193-701.601 embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is mesenchymal progenitor cells.

[0198] In some embodiments, the first subpopulation of fetal cells is early gestation trophoblasts and the second subpopulation of fetal cells is vascular endothelial cells. In some embodiments, the first subpopulation of fetal cells is later gestation trophoblasts and the second subpopulation of fetal cells is vascular endothelial cells.

[0199] In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells is megakaryocytes. In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells is stromal cells. In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells are endothelial cells. In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells is lymphocytes. In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells is nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells is erythroblasts and the second subpopulation of fetal cells is vascular endothelial cells.

[0200] In some embodiments, the first subpopulation of fetal cells is megakaryocytes and the second population of fetal cells is stromal cells. In some embodiments, the first subpopulation of fetal cells is megakaryocytes and the second population of fetal cells is endothelial cells. In some embodiments, the first subpopulation of fetal cells is megakaryocytes and the second population of fetal cells is lymphocytes. In some embodiments, the first subpopulation of fetal cells is megakaryocytes and the second subpopulation of fetal cells is nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells is megakaryocytes and the second subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells is megakaryocytes and the second subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells is megakaryocytes and the second population of fetal cells is vascular endothelial cells.

[0201] In some embodiments, the first subpopulation of fetal cells is stromal cells and the second subpopulation of fetal cells is endothelial cells. In some embodiments, the first subpopulation of fetal cells is stromal cells and the second subpopulation of fetal cells is lymphocytes. In some embodiments, the first subpopulation of fetal cells is stromal cells and theWSGR Docket No. 62193-701.601 second subpopulation of fetal cells is nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells is stromal cells and the second subpopu lation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells is stromal cells and the second subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells is stromal cells and the second subpopulation of fetal cells is vascular endothelial cells.

[0202] In some embodiments, the first subpopulation of fetal cells is endothelial cells and the second subpopulation of fetal cells is lymphocytes. In some embodiments, the first subpopulation of fetal cells is endothelial cells and the second subpopulation of fetal cells is nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells is endothelial cells and the second subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells is endothelial cells and the second subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells is endothelial cells and the second subpopulation of fetal cells is vascular endothelial cells.

[0203] In some embodiments, the first subpopulation of fetal cells is lymphocytes and the second subpopulation of fetal cells is nucleated red blood cells. In some embodiments, the first subpopulation of fetal cells is lymphocytes and the second subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells is lymphocytes and the second subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells is lymphocytes and the second subpopulation of fetal cells is vascular endothelial cells.

[0204] In some embodiments, the first subpopulation of fetal cells is nucleated red blood cells and the second subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation of fetal cells is nucleated red blood cells and the second subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells is nucleated red blood cells and the second subpopulation of fetal cells is vascular endothelial cells.

[0205] In some embodiments, the first subpopulation of fetal cells is hematopoietic progenitor cells and the second subpopulation of fetal cells is mesenchymal progenitor cells. In some embodiments, the first subpopulation of fetal cells is hematopoietic progenitor cells and the second subpopulation of fetal cells is vascular endothelial cells. In some embodiments, the first subpopulation of fetal cells is mesenchymal progenitor cells and the second subpopulation of fetal cells is hematopoietic progenitor cells. In some embodiments, the first subpopulation ofWSGR Docket No. 62193-701.601 fetal cells is mesenchymal progenitor cells and the second subpopulation of fetal cells is vascular endothelial cells.

[0206] In some embodiments, the method further comprises isolating the subpopulation of fetal cells from the maternal biological sample prior to sequencing. In some embodiments, the subpopulation of fetal cells is isolated from the maternal biological sample. In some embodiments, the subpopulation of fetal cells is isolated by contacting the maternal biological sample with a first isolation agent. In some embodiments, the maternal biological sample is contacted with one or more isolation agents. In some embodiments, the one or more isolation agents has a first isolation agent. In some embodiments, the first isolation agent couples to a first biomarker in the subpopulation of fetal cells.

[0207] In some embodiments, the method further comprises isolating the subpopulation of maternal cells from the maternal biological sample prior to sequencing. In some embodiments, the subpopulation of maternal cells is isolated from the maternal biological sample. In some embodiments, the subpopulation of maternal cells is isolated by contacting the maternal biological sample with one or more isolation agents. In some embodiments, the isolation agents include a second isolation agent. In some embodiments, the second isolation agent couples to a second biomarker in the subpopulation of maternal cells.

[0208] In some embodiments, the method further comprises incubating the maternal biological sample after the contacting. In some embodiments, subsequent to the contacting, the maternal biological sample is incubated with one or more isolation agents specific for one or more fetal biomarkers. In some embodiments, the maternal biological sample is incubated with at least the one or more isolation agents specific for one or more fetal biomarkers from about 0 hours to about 48 hours.

[0209] In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more fetal biomarkers from about 0 hour to about 1 hour, from about 1 hour to about 2 hour, from about 2 hours to about 3 hours, from about 3 hours to about 4 hours, from about 4 hours to about 5 hours, from about 5 hours to about 6 hours, from about 6 hours to about 7 hours, from about 7 hours to about 8 hours, from about 8 hours to about 9 hours, from about 9 hours to about 10 hours, from about 10 hours to about 11 hours, from about 11 hours to about 12 hours, from about 12 hours to about 13 hours, from about 13 hours to about 14 hours, from about 14 hours to about 15 hours, from about 15 hours to about 16 hours, from about 16 hours to about 17 hours, from about 17 hours to about 18 hours, from about 18 hours to about 19 hours, from about 19 hours to about 20 hours, from about 20 hours to about 21 hours, from about 21 hours to about 22 hours, from about 22 hours to about 23 hours, fromWSGR Docket No. 62193-701.601 about 23 hours to about 24 hours, from about 24 hours to about 26 hours, from about 26 hours to about 28 hours, from about 28 hours to about 30 hours, from about 30 hours to about 32 hours, from about 32 hours to about 34 hours, from about 34 hours to about 36 hours, from about 36 hours to about 38 hours, from about 38 hours to about 40 hours, from about 40 hours to about 42 hours, from about 42 hours to about 44 hours, from about 44 hours to about 46 hours, from about 46 hours to about 48 hours.

[0210] In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more fetal biomarkers for at least about 0 minutes, at leas t about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes, at least about 25 minutes, at least about 30 minutes, at least about 35 minutes, at least about 40 minutes, at least about 45 minutes, at least about 50 minutes, at least about 55 minutes, at least about 1 hour, at least about 1.25 hours, at least about 1.5 hours, at least about 1.75 hours, at least about 2 hours, at least about 2.25 hours, at least about 2.5 hours, at least about 2.75 hours, at least about 3 hours, at least about 3.25 hours, at least about 3.5 hours, at least about 3.75 hours, at least about 4 hours, at least about 4.25 hours, at least about 4.5 hours, at least about 4.75 hours, at least about 5 hours, at least about 5.25 hours, at least abou t 5.5 hours, at least about 5.75 hours, at least about 6 hours, at least about 6.25 hours, at least about 6.5 hours, at least about 6.75 hours, at least about 7 hours, at least about 7.25 hours, at least about 7.5 hours, at least about 7.75 hours, at least about 8 hours, at least about 8.25 hours, at least about 8.5 hours, at least about 8.75 hours, at least about 9 hours, at least about 9.25 hours, at least about 9.5 hours, at least about 9.75 hours, at least about 10 hours, at least about 10.25 hours, at least about 10.5 hours, at least about 10.75 hours, at least about 11 hours, at least about 11.25 hours, at least about 11.5 hours, at least about 11.75 hours, at least about 12 hours, at least about 13 hours, at least about 14 hours, at least about 15 hours, at least about 16 hours, at least about 17 hours, at least about 18 hours, at least about 19 hours, at least about 20 hours, at least about 21 hours, at least about 22 hours, at least about 23 hours, at least about 24 hours, at least about 25 hours, at least about 26 hours, at least about 27 hours, at least about 28 hours, at least about 29 hours, at least about 30 hours, at least about 31 hours, at least about 32 hours, at least about 33 hours, at least about 34 hours, at least about 35 hours, at least about 36 hours, at least about 37 hours, at least about 38 hours, at least about 39 hours, at least about 40 hours, at least about 41 hours, at least about 42 hours, at least about 43 hours, at least about 44 hours, at least about 45 hours, at least about 46 hours, at least about 47 hours, at least about 48 hours, or more.

[0211] In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more fetal biomarkers for at most about 48 hours, at mostWSGR Docket No. 62193-701.601 about 47 hours, at most about 46 hours, at most about 45 hours, at most about 44 hours, at most about 43 hours, at most about 42 hours, at most about 41 hours, at most about 40 hours, at most about 39 hours, at most about 38 hours, at most about 37 hours, at most about 36 hours, at most about 35 hours, at most about 34 hours, at most about 33 hours, at most about 32 hours, at most about 31 hours, at most about 30 hours, at most about 29 hours, at most about 28 hours, at most about 27 hours, at most about 26 hours, at most about 25 hours, at most about 24 hours, at most about 23 hours, at most about 22 hours, at most about 21 hours, at most about 20 hours, at most about 19 hours, at most about 18 hours, at most about 17 hours, at most about 16 hours, at most about 15 hours, at most about 14 hours, at most about 13 hours, at most about 12 hours, at most about 11.75 hours, at most about 11.5 hours, at most about 11.25 hours, at most about 11 hours, at most about 10.75 hours, at most about 10.5 hours, at most about 10.25 hours, at most about 10 hours, at most about 9.75 hours, at most about 9.5 hours, at most about 9.25 hours, at most about 9 hours, at most about 8.75 hours, at most about 8.5 hours, at most about 8.25 hours, at most about 8 hours, at most about 7.75 hours, at most about 7.5 hours, at most about 7.25 hours, at most about 7 hours, at most about 6.75 hours, at most about 6.5 hours, at most about 6.25 hours, at most about 6 hours, at most about 5.75 hours, at most about 5.5 hours, at most about 5.25 hours, at most about 5 hours, at most about 4.75 hours, at most about 4.5 hours, at most about 4.25 hours, at most about 4 hours, at most about 3.75 hours, at most about 3.5 hours, at most about 3.25 hours, at most about 3 hours, at most about 2.75 hours, at most about 2.5 hours, at most about 2.25 hours, at most about 2 hours, at most about 1.75 hours, at most about 1.5 hours, at most about 1.25 hours, at most about 1 hour, at most about 55 minutes, at most about 50 minutes, at most about 45 minutes, at most about 40 minutes, at most about 35 minutes, at most about 30 minutes, at most about 25 minutes, at most about 20 minutes, at most about 15 minutes, at most about 10 minutes, at most about 5 minutes, at most about 0 minutes, or less.

[0212] In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more fetal biomarkers for about 0 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 1 hour, about 1.25 hours, about 1.5 hours, about 1.75 hours, about 2 hours, about 2.25 hours, about 2.5 hours, about 2.75 hours, about 3 hours, about 3.25 hours, about 3.5 hours, about 3.75 hours, about 4 hours, about 4.25 hours, about 4.5 hours, about 4.75 hours, about 5 hours, about 5.25 hours, about 5.5 hours, about 5.75 hours, about 6 hours, about 6.25 hours about 6.5 hours, about 6.75 hours, about 7 hours, about 7.25 hours, about 7.5 h ours, about 7.75 hours, about 8 hours, about 8.25 hours, about 8.5 hours, about 8.75 hours, about 9 hours, about 9.25WSGR Docket No. 62193-701.601 hours, about 9.5 hours, about 9.75 hours, about 10 hours, about 10.25 hours, about 10.5 hours, about 10.75 hours, about 11 hours, about 11.25 hours, about 11.5 hours, about 11.75 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 25 hours, about 26 hours, about 27 hours, about 28 hours, about 29 hours, about 30 hours, about 31 hours, about 32 hours, about 33 hours, about 34 hours, about 35 hours, about 36 hours, about 37 hours, about 38 hours, about 39 hours, about 40 hours, about 41 hours, about 42 hours, about 43 hours, about 44 hours, about 45 hours, about 46 hours, about 47, or about 48 hours.

[0213] In some embodiments, the maternal biological sample is incubated with the at least one isolation agent specific for one or more fetal biomarkers at a temperature between about 20°C to about 70°C. In some embodiments, the maternal biological sample is incubated with the at least one isolation agent specific for one or more fetal biomarkers at a temperature of at least about 20°C, at least about 21°C, at least about 22°C, at least about 23°C, at least about 24°C, at least about 25°C, at least about 26°C, at least about 27°C, at least about 28°C, at least about 29°C, at least about 30°C, at least about 31°C, at least about 32°C, at least about 33°C, at least about 34°C, at least about 35°C, at least about 36°C, at least about 37°C, at least about 38°C, at least about 39°C, at least about 40°C, at least about 41°C, at least about 42°C, at least about 43°C, at least about 44°C, at least about 45°C, at least about 46°C, at least about 47°C, at least about 48°C, at least about 49°C, at least about 50°C, at least about 51°C, at least about 52°C, at least about 53°C, at least about 54°C, at least about 55°C, at least about 56°C, at least about 57°C, at least about 58°C, at least about 59°C, at least about 60°C, at least about 61°C, at least about 62°C, at least about 63°C, at least about 64°C, at least about 65°C, at least about 66°C, at least about 67°C, at least about 68°C, at least about 69°C, at least about 70°C, or more. In some embodiments, the maternal biological sample is incubated with the at least one isolation agent specific for one or more fetal biomarkers at a temperature of at most about 70°C, at most about 69°C, at most about 68°C, at most about 67°C, at most about 66°C, at most about 65°C, at most about 64°C, at most about 63°C, at most about 62°C, at most about 61°C, most about 60°C, at most about 59°C, at most about 58°C, at most about 57°C, at most about 56°C, at most about 55°C, at most about 54°C, at most about 53°C, at most about 52°C, at most about 51°C, at most about 50°C, at most about 49°C, at most about 48°C, at most about 47°C, at most about 46°C, at most about 45°C, at most about 44°C, at most about 43°C, at most about 42°C, at most about 41°C, at most about 40°C, at most about 39°C, at most about 38°C, at most about 37°C, at most about 36°C, at most about 35°C, at most about 34°C, at most about 33°C, at most about 32°C, atWSGR Docket No. 62193-701.601 most about 31°C, at most about 30°C, at most about 29°C, at most about 28°C, at most about 27°C, at most about 26°C, at most about 25°C, at most about 24°C, at most about 23°C, at most about 22°C, at most about 21°C, at most about 20°C, or less. In some embodiments, the maternal biological sample is incubated with the at least one isolation agent specific to one or more fetal biomarkers at a temperature of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, about 40°C, about 41°C, about 42°C, about 43°C, about 44°C, about 45°C, about 46°C, about 47°C, about 48°C, about 49°C, about 50°C, about 51°C, about 52°C, about 53°C, about 54°C, about 55°C, about 56°C, about 57°C, about 58°C, about 59°C, about 60°C, about 61°C, about 62°C, about 63°C, about 64°C, about 65°C, about 66°C, about 67°C, about 68°C, about 69°C, or about 70°C.

[0214] In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more fetal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more maternal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more fetal biomarkers, then subsequently incubated with one or more isolation agents that couples to one or more maternal biomarkers. In some embodiments, that maternal biological sample is incubated with one or more isolation agents that couples to one or more maternal biomarkers, then subsequently incubated with one or more isolation agents that couples to one or more fetal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more fetal biomarkers, then subsequently incubated with another one or more isolation agents that couples to another one or more fetal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more maternal biomarkers, then incubated with one or more isolation agents that couples to one or more fetal biomarkers, and then incubated with another one or more isolation agents that couples to another one or more fetal biomarkers.

[0215] In some embodiments, the first isolation agent couples to a first biomarker. In some embodiments, the first biomarker is one or more fetal biomarkers. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to an extracellular biomarker. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to an extracellular protein. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a cell surface protein. In someWSGR Docket No. 62193-701.601 embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a soluble cell surface protein. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a cell-surface receptor. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a transmembrane protein. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a transmembrane channel protein.

[0216] In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to an intracellular biomarker. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to an intracellular protein. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to an enzyme. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a phosphoprotein. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a growth factor. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a structural protein.

[0217] In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a nucleic acid biomarker. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a DNA biomarker. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to an RNA biomarker. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to mRNA. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to rRNA. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to lncRNA. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to miRNA. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to siRNA.

[0218] In some embodiments, the first isolation agent couples to one or more fetal biomarkers. In some embodiments, the first biomarker is one or more fetal biomarkers. In some embodiments, the first biomarker is a Sct biomarker, a H19 biomarker, a Tfpi biomarker, or a Fthl17a biomarker. In some embodiments, the first biomarker is a Sct biomarker. In some embodiments, the first biomarker is a H19 biomarker. In some embodiments, the first biomarker is a Tfpi biomarker. In some embodiments, the first biomarker is a Fthl17a biomarker.

[0219] In some embodiments, the method further comprises isolating the subpopulation of fetal cells from the maternal biological sample prior to sequencing. In some embodiments, theWSGR Docket No. 62193-701.601 subpopulation of fetal cells is isolated from the maternal biological sample. In some embodiments, the subpopulation of fetal cells is isolated by contacting the maternal biological sample with one or more isolation agents specific for one or more maternal biomarkers. In some embodiments, the maternal biological sample is contacted with one or more isolation agents specific for one or more maternal biomarkers. In some embodiments, the one or more isolation agents specific for one or more maternal biomarkers to a second biomarker in the subpopulation of maternal cells.

[0220] In some embodiments, the method further comprises isolating the subpopulation of maternal cells from the maternal biological sample prior to sequencing. In some embodiments, the subpopulation of maternal cells is isolated from the maternal biological sample. In some embodiments, the subpopulation of maternal cells is isolated by contacting the maternal biological sample with a one or more isolation agents specific for one or more fetal biomarkers. In some embodiments, the one or more isolation agents specific for one or more fetal biomarkers couples to a first biomarker in the subpopulation of fetal cells.

[0221] In some embodiments, the method further comprises incubating the maternal biological sample after the contacting. In some embodiments, subsequent to the contacting, the maternal biological sample is incubated with the one or more isolation agents specific for one or more maternal biomarkers. In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more maternal biomarkers from about 0 hours to about 48 hours.

[0222] In some embodiments, the maternal biological sample is incubated with the second isolation agent from about 0 hour to about 1 hour, from about 1 hour to about 2 hour, from about 2 hours to about 3 hours, from about 3 hours to about 4 hours, from about 4 hours to about 5 hours, from about 5 hours to about 6 hours, from about 6 hours to about 7 hours, from about 7 hours to about 8 hours, from about 8 hours to about 9 hours, from about 9 hours to about 10 hours, from about 10 hours to about 11 hours, from about 11 hours to about 12 hours, from about 12 hours to about 13 hours, from about 13 hours to about 14 hours, from about 14 hours to about 15 hours, from about 15 hours to about 16 hours, from about 16 hours to about 17 hours, from about 17 hours to about 18 hours, from about 18 hours to about 19 hours , from about 19 hours to about 20 hours, from about 20 hours to about 21 hours, from about 21 hours to about 22 hours, from about 22 hours to about 23 hours, from about 23 hours to about 24 hours, from about 24 hours to about 26 hours, from about 26 hours to about 28 hours, from about 28 hours to about 30 hours, from about 30 hours to about 32 hours, from about 32 hours to about 34 hours, from about 34 hours to about 36 hours, from about 36 hours to about 38 hours, from about 38WSGR Docket No. 62193-701.601 hours to about 40 hours, from about 40 hours to about 42 hours, from about 42 hours to about 44 hours, from about 44 hours to about 46 hours, from about 46 hours to about 48 hours.

[0223] In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more maternal biomarkers for at least about 0 minutes, at least about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes, at least about 25 minutes, at least about 30 minutes, at least about 35 minutes, at least about 40 minutes, at least about 45 minutes, at least about 50 minutes, at least about 55 minutes, at least about 1 hour, at least about 1.25 hours, at least about 1.5 hours, at least about 1.75 hours, at least about 2 hours, at least about 2.25 hours, at least about 2.5 hours, at least about 2.75 hours, at least about 3 hours, at least about 3.25 hours, at least about 3.5 hours, at least about 3.75 hours, at least about 4 hours, at least about 4.25 hours, at least about 4.5 hours, at least about 4.75 hours, at least about 5 hours, at least about 5.25 hours, at least about 5.5 hours, at least about 5.75 hours, at least about 6 hours, at least about 6.25 hours, at least about 6.5 hours, at least about 6.75 hours, at least about 7 hours, at least about 7.25 hours, at least about 7.5 hours, at least about 7.75 hours, at least about 8 hours, at least about 8.25 hours, at least about 8.5 hours, at least about 8.75 hours, at least about 9 hours, at least about 9.25 hours, at least about 9.5 hours, at least about 9.75 hours, at least about 10 hours, at least about 10.25 hours, at least about 10.5 hours, at least about 10.75 hours, at least about 11 hours, at least about 11.25 hours, at least about 11.5 hours, at least about 11.75 hours, at least about 12 hours, at least about 13 hours, at least about 14 hours, at least about 15 hours, at least about 16 hours, at least about 17 hours, at least about 18 hours, at least about 19 hours, at least about 20 hours, at least about 21 hours, at least about 22 hours, at least about 23 hours, at least about 24 hours, at least about 25 hours, at least about 26 hours, at least about 27 hours, at least about 28 hours, at least about 29 hours, at least about 30 hours, at least about 31 hours, at least about 32 hours, at least about 33 hours, at least about 34 hours, at least about 35 hours, at least about 36 hours, at least about 37 hours, at least about 38 hours, at least about 39 hours, at least about 40 hours, at least about 41 hours, at least about 42 hours, at least about 43 hours, at least about 44 hours, at least about 45 hours, at least about 46 hours, at least about 47 hours, at least about 48 hours, or more.

[0224] In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more maternal biomarkers for at most about 48 hours, at most about 47 hours, at most about 46 hours, at most about 45 hours, at most about 44 hours, at most about 43 hours, at most about 42 hours, at most about 41 hours, at most about 40 hours, at most about 39 hours, at most about 38 hours, at most about 37 hours, at most about 36 hours, at most about 35 hours, at most about 34 hours, at most about 33 hours, at most about 32 hours, at mostWSGR Docket No. 62193-701.601 about 31 hours, at most about 30 hours, at most about 29 hours, at most about 28 hours, at most about 27 hours, at most about 26 hours, at most about 25 hours, at most about 24 hours, at most about 23 hours, at most about 22 hours, at most about 21 hours, at most about 20 hours, at most about 19 hours, at most about 18 hours, at most about 17 hours, at most about 16 hours, at most about 15 hours, at most about 14 hours, at most about 13 hours, at most about 12 hours, at most about 11.75 hours, at most about 11.5 hours, at most about 11.25 hours, at most about 11 hours, at most about 10.75 hours, at most about 10.5 hours, at most about 10.25 hours, at most about 10 hours, at most about 9.75 hours, at most about 9.5 hours, at most about 9.25 hours, at most about 9 hours, at most about 8.75 hours, at most about 8.5 hours, at most about 8.25 hours, at most about 8 hours, at most about 7.75 hours, at most about 7.5 hours, at most about 7.25 hours, at most about 7 hours, at most about 6.75 hours, at most about 6.5 hours, at most about 6.25 hours, at most about 6 hours, at most about 5.75 hours, at most about 5.5 hours, at most about 5.25 hours, at most about 5 hours, at most about 4.75 hours, at most about 4.5 hours, at most about 4.25 hours, at most about 4 hours, at most about 3.75 hours, at most about 3.5 ho urs, at most about 3.25 hours, at most about 3 hours, at most about 2.75 hours, at most about 2.5 hours, at most about 2.25 hours, at most about 2 hours, at most about 1.75 hours, at most about 1.5 hours, at most about 1.25 hours, at most about 1 hour, at most about 55 minutes, at most about 50 minutes, at most about 45 minutes, at most about 40 minutes, at most about 35 minutes, at most about 30 minutes, at most about 25 minutes, at most about 20 minutes, at most about 15 minutes, at most about 10 minutes, at most about 5 minutes, at most about 0 minutes, or less.

[0225] In some embodiments, the maternal biological sample is incubated with the one or more isolation agents specific for one or more maternal biomarkers for about 0 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 1 hour, about 1.25 hours, about 1.5 hours, about 1.75 hours, about 2 hours, abou t 2.25 hours, about 2.5 hours, about 2.75 hours, about 3 hours, about 3.25 hours, about 3.5 hours, about 3.75 hours, about 4 hours, about 4.25 hours, about 4.5 hours, about 4.75 hours, about 5 hours, about 5.25 hours, about 5.5 hours, about 5.75 hours, abo ut 6 hours, about 6.25 hours about 6.5 hours, about 6.75 hours, about 7 hours, about 7.25 hours, about 7.5 hours, about 7.75 hours, about 8 hours, about 8.25 hours, about 8.5 hours, about 8.75 hours, about 9 hours, about 9.25 hours, about 9.5 hours, about 9.75 hours, about 10 hours, about 10.25 hours, about 10.5 hours, about 10.75 hours, about 11 hours, about 11.25 hours, about 11.5 hours, about 11.75 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23WSGR Docket No. 62193-701.601 hours, about 24 hours, about 25 hours, about 26 hours, about 27 hours, about 28 hours, about 29 hours, about 30 hours, about 31 hours, about 32 hours, about 33 hours, abou t 34 hours, about 35 hours, about 36 hours, about 37 hours, about 38 hours, about 39 hours, about 40 hours, about 41 hours, about 42 hours, about 43 hours, about 44 hours, about 45 hours, about 46 hours, about 47, or about 48 hours.

[0226] In some embodiments, the maternal biological sample is incubated with the at least one isolation agent specific for one or more maternal biomarkers at a temperature between about 20°C to about 70°C. In some embodiments, the maternal biological sample is incubated with the at least one isolation agent specific for one or more maternal biomarkers at a temperature of at least about 20°C, at least about 21°C, at least about 22°C, at least about 23°C, at least about 24°C, at least about 25°C, at least about 26°C, at least about 27°C, at least about 28°C, at least about 29°C, at least about 30°C, at least about 31°C, at least about 32°C, at least about 33°C, at least about 34°C, at least about 35°C, at least about 36°C, at least about 37°C, at least about 38°C, at least about 39°C, at least about 40°C, at least about 41°C, at least about 42°C, at least about 43°C, at least about 44°C, at least about 45°C, at least about 46°C, at least about 47°C, at least about 48°C, at least about 49°C, at least about 50°C, at least about 51°C, at least about 52°C, at least about 53°C, at least about 54°C, at least about 55°C, at least about 56°C, at least about 57°C, at least about 58°C, at least about 59°C, at least about 60°C, at least about 61°C, at least about 62°C, at least about 63°C, at least about 64°C, at least about 65°C, at least about 66°C, at least about 67°C, at least about 68°C, at least about 69°C, at least about 70°C, or more. In some embodiments, the maternal biological sample is incubated with the at least one isolation agent specific for one or more maternal biomarkers at a temperature of at most about 70°C, at most about 69°C, at most about 68°C, at most about 67°C, at most about 66°C, at most about 65°C, at most about 64°C, at most about 63°C, at most about 62°C, at most about 61°C, most about 60°C, at most about 59°C, at most about 58°C, at most about 57°C, at most about 56°C, at most about 55°C, at most about 54°C, at most about 53°C, at most about 52°C, at most about 51°C, at most about 50°C, at most about 49°C, at most about 48°C, at most about 47°C, at most about 46°C, at most about 45°C, at most about 44°C, at most about 43°C, at most about 42°C, at most about 41°C, at most about 40°C, at most about 39°C, at most about 38°C, at most about 37°C, at most about 36°C, at most about 35°C, at most about 34°C, at most about 33°C, at most about 32°C, at most about 31°C, at most about 30°C, at most about 29°C, at most about 28°C, at most about 27°C, at most about 26°C, at most about 25°C, at most about 24°C, at most about 23°C, at most about 22°C, at most about 21°C, at most about 20°C, or less. In some embodiments, the maternal biological sample is incubated with the at least one isolation agentWSGR Docket No. 62193-701.601 specific for one or more maternal biomarkers at a temperature of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, about 40°C, about 41°C, about 42°C, about 43°C, about 44°C, about 45°C, about 46°C, about 47°C, about 48°C, about 49°C, about 50°C, about 51°C, about 52°C, about 53°C, about 54°C, about 55°C, about 56°C, about 57°C, about 58°C, about 59°C, about 60°C, about 61°C, about 62°C, about 63°C, about 64°C, about 65°C, about 66°C, about 67°C, about 68°C, about 69°C, or about 70°C.

[0227] In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more fetal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more maternal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more fetal biomarkers, then subsequently incubated with one or more isolation agents that couples to one or more maternal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more maternal biomarkers, then subsequently incubated with one or more isolation agents that couples to one or more fetal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more fetal biomarkers, then subsequently incubated with another one or most isolation agents that couples to another one or more fetal biomarkers. In some embodiments, the maternal biological sample is incubated with one or more isolation agents that couples to one or more maternal biomarkers, then incubated with one or more isolation agents that couples to one or more fetal biomarkers, and then incubated with another one or more isolation agents that couples to another one or more fetal biomarkers.

[0228] In some embodiments, the maternal biological sample undergoes at least one round of enrichment to enrich / isolate fetal cells from the maternal blood sample. In some embodiments, the maternal biological sample undergoes between about one round of enrichment and about ten rounds of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes at least about one round, at least about two rounds, at least about three rounds, at least about four rounds, at least about five rounds, at least about six rounds, at least about seven rounds, at least about eight rounds, at least about nine rounds, or at least about ten rounds or more of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes at most about ten rounds, at most about nine rounds, at most about eight rounds, at most aboutWSGR Docket No. 62193-701.601 seven rounds, at most about six rounds, at most about five rounds, at most about four rounds, at most about three rounds, at most about two rounds, at most about one round, or less of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes about one round, about two rounds, about three rounds, about four rounds, about five rounds, about six rounds, about seven rounds, about eight rounds, about nine rounds, or about ten rounds of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes about one round of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes about two rounds of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes about three rounds of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes about four rounds of enrichment to enrich / isolate fetal cells from the maternal biological sample. In some embodiments, the maternal biological sample undergoes about five rounds of enrichment to enrich / isolate fetal cells from the maternal biological sample.

[0229] In some embodiments, the at least one round of enrichment enriches for at least one fetal biomarker. In some embodiments, the at least one round of enrichment enriches for at least one maternal biomarker. In some embodiments, the at least one round of enrichment enriches for maternal biomarkers, subsequently followed by enriching for at least one fetal biomarker. In some embodiments, the at least one round of enrichment enriches for at least one maternal biomarkers, followed by enriching for at least one fetal biomarkers, and then followed by enriching for at least another fetal biomarker. In some embodiments, the at least one round of enrichment enriches for fetal biomarkers, subsequently followed by enriching for at least another fetal biomarker.

[0230] In some embodiments, the at least one fetal biomarker is about one fetal biomarker disclosed herein. In some embodiments, the at least one fetal biomarker is at least two of more fetal biomarkers disclosed herein. In some embodiments, the at least one feta l biomarker is between about two fetal biomarkers to about ten fetal biomarkers. In some embodiments, the at least one fetal biomarker is at least one about fetal biomarker, at least about two fetal biomarkers, at least about three fetal biomarkers, at least about four fetal biomarkers, at least about five fetal biomarkers, at least about six fetal biomarkers, at least about seven fetal biomarkers, at least about eight fetal biomarkers, at least about nine fetal biomarkers, or at least about ten fetal biomarkers. In some embodiments, the at least one fetal biomarker is at mostWSGR Docket No. 62193-701.601 about ten fetal biomarkers, at most about nine fetal biomarkers, at most about eight fetal biomarkers, at most about seven fetal biomarkers, at most about six fetal biomarkers, at most about five fetal biomarkers, at most about four fetal biomarkers, at most about three fetal biomarkers, at most about two fetal biomarkers, at most about one fetal biomarker, or less. In some embodiments, the at least one fetal biomarker is about one fetal biomarker, about two fetal biomarkers, about three fetal biomarkers, about four fetal biomarkers, about five fetal biomarkers, about six fetal biomarkers, about seven fetal biomarkers, about eight fetal biomarkers, about nine fetal biomarkers, or about ten fetal biomarkers. In some embodiments, the at least one fetal biomarker is about two fetal biomarkers. In some embodiments, the at least one fetal biomarker is about three fetal biomarkers. In some embodiments, the at least one fetal biomarker is about four fetal biomarkers. In some embodiments, the at least one fetal biomarker is about five fetal biomarkers. In some embodiments, the at least one fetal biomarker is any combination of fetal biomarkers disclosed within the present disclosure.

[0231] In some embodiments, the at least one maternal biomarker is about one maternal biomarker disclosed herein. In some embodiments, the at least one maternal biomarker is at least two of more maternal biomarkers disclosed herein. In some embodiments, the at least one maternal biomarker is between about two maternal biomarkers to about ten maternal biomarkers. In some embodiments, the at least one maternal biomarker is at least one about maternal biomarker, at least about two maternal biomarkers, at least about three maternal biomarkers, at least about four maternal biomarkers, at least about five maternal biomarkers, at least about six maternal biomarkers, at least about seven maternal biomarkers, at least about eight maternal biomarkers, at least about nine maternal biomarkers, or at least about ten maternal biomarkers. In some embodiments, the at least one maternal biomarker is at most about ten maternal biomarkers, at most about nine maternal biomarkers, at most about eight maternal biomarkers, at most about seven maternal biomarkers, at most about six maternal biomarkers, at most about five maternal biomarkers, at most about four maternal biomarkers, at most about three maternal biomarkers, at most about two maternal biomarkers, at most about one maternal biomarker, or less. In some embodiments, the at least one maternal biomarker is about one maternal biomarker, about two maternal biomarkers, about three maternal biomarkers, about four maternal biomarkers, about five maternal biomarkers, about six maternal biomarkers, about seven maternal biomarkers, about eight maternal biomarkers, about nine maternal biomarkers, or about ten maternal biomarkers. In some embodiments, the at least one maternal biomarker is about two maternal biomarkers. In some embodiments, the at least one maternal biomarker is about three maternal biomarkers. In some embodiments, the at least one maternal biomarker isWSGR Docket No. 62193-701.601 about four maternal biomarkers. In some embodiments, the at least one maternal biomarker is about five maternal biomarkers. In some embodiments, the at least one maternal biomarker is any combination of maternal biomarkers disclosed within the present disclosure.

[0232] In some embodiments, the one or more isolation agents specific for one or more maternal biomarkers couples to a second biomarker. In some embodiments, the second biomarker is a maternal biomarker. In some embodiments, the isolation agent specific for a maternal biomarker couples to an extracellular biomarker. In some embodiments, the isolation agent specific for a maternal biomarker couples to an extracellular protein. In some embodiments, the isolation agent specific for a maternal biomarker couples to a cell surface protein. In some embodiments, the isolation agent specific for a maternal biomarker couples to a soluble cell surface p rotein. In some embodiments, the isolation agent specific for a maternal biomarker couples to a cell- surface receptor. In some embodiments, the isolation agent specific for a maternal biomarker couples to a transmembrane protein. In some embodiments, the isolation agent specific for a maternal biomarker couples to a transmembrane channel protein.

[0233] In some embodiments, the isolation agent specific for a maternal biomarker couples to an intracellular biomarker. In some embodiments, the isolation agent specific for a maternal biomarker couples to an intracellular protein. In some embodiments, the isolation agent specific for a maternal biomarker couples to an enzyme. In some embodiments, the isolation agents specific for a maternal biomarker couples to a phosphoprotein. In some embodiments, the isolation agent specific for a maternal biomarker couples to a growth factor. In some embodiments, the isolation agent specific for a maternal biomarker couples to a structural protein.

[0234] In some embodiments, the isolation agent specific for a maternal biomarker couples to a nucleic acid biomarker. In some embodiments, the isolation agent specific for a maternal biomarker couples to a DNA biomarker. In some embodiments, the isolation agent specific for a maternal biomarker couples to an RNA biomarker. In some embodiments, the isolation agent specific for a maternal biomarker couples to mRNA. In some embodiments, the isolation agent specific for a maternal biomarker couples to rRNA. In some embodiments, the isolation agent specific for a maternal biomarker couples to lncRNA. In some embodiments, the isolation agent specific for a maternal biomarker couples to miRNA. In some embodiments, the second isolation agent couples to siRNA.

[0235] In some embodiments, the second isolation agent couples to a maternal biomarkers. In some embodiments, the second biomarker is one or more maternal biomarkers. In some embodiments, the second biomarker is a Coro1a biomarker, a Cd52 biomarker, a CybaWSGR Docket No. 62193-701.601 biomarker, a Rac2 biomarker, a Laptm5 biomarker, or a Ptprc biomarker. In some embodiments, the second biomarker is a Coro1a biomarker. In some embodiments, the second biomarker is a Cd52 biomarker. In some embodiments, the second biomarker is a Cyba biomarker. In some embodiments, the second biomarker is a Rac2 biomarker. In some embodiments, the second biomarker is a Laptm5 biomarker. In some embodiments, the second biomarker is a Ptprc biomarker.

[0236] After obtaining the subpopulation of fetal cells and / or the subpopulation of maternal cells, the one or more isolation agents bound to the one or more biomarkers can be removed. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by one or more washing operations. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifugation. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifugation of the subpopulation of fetal cells and / or the subpopulation of maternal cells from between 1200 revolutions per minute (rpm) to about 1800 rpm. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifugation of the subpopulation of fetal cells and / or the subpopulation of maternal cells at a rate of at least about 1200 rpm, at least about 1250 rpm, at least about 1300 rpm, at least about 1350 rpm, at least about 1400 rpm, at least about 1450 rpm, at least about 1500 rpm, at least about 1550 rpm, at least about 1600 rpm, at least about 1650 rpm, at least about 1700 rpm, at least about 1750 rpm, at least about 1800 rpm, or more. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifuging the subpopulation of fetal cells and / or the subpopulation of maternal cells at a rate of at most 1800 rpm, at most about 1750 rpm, at most about 1700 rpm, at most about 1650 rpm, at most about 1600 rpm, at most about 1550 rpm, at most about 1500 rpm, at most about 1450 rpm, at most about 1400 rpm, at most about 1350 rpm, at most about 1300 rpm, at most about 1250 rpm, at most about 1200 rpm, or less. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifugation of the subpopulation of fetal cells and / or the subpopulation of maternal cells at about 1200 rpm, about 1250 rpm, about 1300 rpm, about 1350 rpm, about 1400 rpm, about 1450 rpm, about 1500 rpm, about 1550 rpm, about 1600 rpm, about 1650 rpm, about 1700 rpm, about 1750 rpm, about 1800 rpm, or more. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifugation of the subpopulation of fetal cells and / or the subpopulation of maternal cells from between about 0 minutes to about 30 minutes. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can beWSGR Docket No. 62193-701.601 removed by centrifugation of the subpopulation of fetal cells and / or the subpopulation of maternal cells for at least about 0 minutes, at least about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes, at least about 25 minutes, at least about 30 minutes, or more. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifugation of the subpopulation of fetal cells and / or the subpopulation of maternal cells for at most about 30 minutes, at most about 25 minutes, at most about 20 minutes, at most about 15 minutes, at most about 10 minutes, at most about 5 minutes, at most about 0 minutes, or less. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by centrifugation of the subpopulation of fetal cells and / or the subpopulation of maternal cells for about 0 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, or about 30 minutes.

[0237] In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or subpopulation of maternal cells to heat. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to heat from between about 0 minutes to about 30 minutes. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to heat for at least about 0 minutes, at least about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes, at least about 25 minutes, at least about 30 minutes, or more. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to heat for at most about 30 minutes, at most about 25 minutes, at most about 20 minutes, at most about 15 minutes, at most about 10 minutes, at most about 5 minutes, at most about 0 minutes, or less. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to heat for about 0 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, or about 30 minutes. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to a temperature of between about 50°C to about 100°C. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to a temperature of at least about 50°C, at least about 55°C, at least about 60°C, at least aboutWSGR Docket No. 62193-701.601 65°C, at least about 70°C, at least about 75°C, at least about 80°C, at least about 85°C, at least about 90°C, at least about 95°C, at least about 100°C, or greater. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to a temperature of at most about 100°C, at most about 95°C, at most about 90°C, at most about 85°C, at most about 80°C, at most about 75°C, at most about 70°C, at most about 65°C, at most about 60°C, at most about 55°C, at most about 50, or less. In some embodiments, the one or more isolation agents bound to the one or more biomarkers can be removed by exposing the subpopulation of fetal cells and / or the subpopulation of maternal cells to a temperature of about 50°C, about 55°C, about 60°C, about 65°C, about 70°C, about 75°C, about 80°C, about 85°C, about 90°C, about 95°C, or about 100°C.

[0238] After obtaining the subpopulation of fetal cells and / or the subpopulation of maternal cells, the fetal cells and / or maternal cells can be processed to generate sequencing data of the genome of the fetal cells. Processing the biological sample obtained from the subject can comprise (i) subjecting the CFCs to conditions that are sufficient to isolate, enrich, or extract a plurality of fetal cell genetic material, and (ii) sequencing the plurality of fetal cell genetic material to generate a plurality of sequencing reads. Processing the biological sample obtained from the subject can comprise (i) subjecting the maternal cells to conditions that are sufficient to isolate, enrich, or extract a plurality of maternal cell genetic material, and (ii) sequencing the plurality of maternal cell genetic material to generate a plurality of sequencing reads. In some embodiments, the plurality of sequencing reads is used to determine whether the fetus (or fetuses) has a genetic disorder. The nucleic acid molecules can comprise ribonucleic acid (RNA) or deoxyribonucleic acid (DNA). The nucleic acid molecules can comprise DNA. The nucleic acid molecules can comprise RNA. The nucleic acid molecules (e.g., RNA or DNA) can be extracted from the subpopulation of fetal cells by a variety of methods, such as a DNeasy Blood and Tissue Kit, a PureLink Genomic DNA kit, or a Wizard® HMW DNA Extraction Kit. The extraction method can extract all RNA or DNA molecules from a sample.

[0239] In some embodiments, the genetic material from the subpopulation of fetal cells is genetically analyzed. In some embodiments, genetic material is analyzed via PCR, sequencing, hybridization methods, or any combination thereof. In some embodiments, the genetic material is analyzed via a whole genome scan. In some embodiments, the genetic material is analyzed via short tandem repeat analysis. In some embodiments, the genetic material is analyzed via microsatellite analysis.WSGR Docket No. 62193-701.601

[0240] In some embodiments, the genetic analysis is a hybridization method. In some embodiments, the hybridization method is a restriction fragment length polymorphism (RFLP), a southern blot analysis, a microarray, a fluorescence in situ hybridization (FISH), a comparative genomic hybridization, in situ hybridization, or any combination thereof.

[0241] In some embodiments, the genetic material from the subpopulation of fetal cells is amplified prior to genetic analysis. In some embodiments, the genetic material from the subpopulation of fetal cells is amplified to increase the amount of the genetic material from the subpopulation of fetal cells by between about 0% to about 500%. In some embodiments, the genetic material from the subpopulation of fetal cells is amplified to increase the amount of genetic material from the subpopulation of fetal cell by at least about 0%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 120%, at least about 140%, at least about 160%, at least about 180%, at least about 200%, at least about 250%, at least about 300%, at least about 350%, at least about 400%, at least about 450%, at least about 500%, or more. In some embodiments, the genetic material from the subpopulation of fetal cells is amplified to increase the amount of genetic material from the subpopulation of fetal cell by at most about 500%, at most about 450%, at most about 400%, at most about 350%, at most about 300%, at most about 250%, at most about 200%, at most about 180%, at most about 160%, at most about 140%, at most about 120%, at most about 100%, at most about 90%, at most about 80%, at most about 70%, at most about 60%, at most about 50%, at most about 40%, at most about 30%, at most about 20%, at most about 10%, at most about 0%, or less. In some embodiments, the genetic material from the subpopulation of fetal cells is amplified to increase the amount of genetic material from the subpopulation of fetal cell by about 0%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 120%, about 140%, about 160%, about 180%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, or about 500%.

[0242] some embodiments, the genetic material from the subpopulation of maternal cells is amplified prior to genetic analysis. In some embodiments, the genetic material from the subpopulation of maternal cells is amplified to increase the amount of the genetic material from the subpopulation of maternal cells by between about 0% to about 500%. In some embodiments, the genetic material from the subpopulation of maternal cells is amplified to increase the amount of genetic material from the subpopulation of maternal cell by at least about 0%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, atWSGR Docket No. 62193-701.601 least about 120%, at least about 140%, at least about 160%, at least about 180%, at least about 200%, at least about 250%, at least about 300%, at least about 350%, at least about 400%, at least about 450%, at least about 500%, or more. In some embodiments, the genetic material from the subpopulation of maternal cells is amplified to increase the amount of genetic material from the subpopulation of maternal cell by at most about 500%, at most about 450%, at most about 400%, at most about 350%, at most about 300%, at most about 250%, at most about 200%, at most about 180%, at most about 160%, at most about 140%, at most about 120%, at most about 100%, at most about 90%, at most about 80%, at most about 70%, at most about 60%, at most about 50%, at most about 40%, at most about 30%, at most about 20%, at most about 10%, at most about 0%, or less. In some embodiments, the genetic material from the subpopulation of maternal cells is amplified to increase the amount of genetic material from the subpopulation of maternal cell by about 0%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 120%, about 140%, about 160%, about 180%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, or about 500%.

[0243] In some embodiments, the genetic analysis is sequencing. In some embodiments, the method further comprises sequencing the nucleic acid molecules from the subpopulation of fetal cells. In some embodiments, the nucleic acid molecules from the subpopulation of fetal cells generate a nucleic acid sequence. In some embodiments, the nucleic acid sequence is from the subpopulation of fetal cells. In some embodiments, the method further comprises sequencing the nucleic acid molecules from the subpopulation of maternal cells. In some embodiments, the nucleic acid molecules from the subpopulation of maternal cells generate another nucleic acid sequence. In some embodiments, the other nucleic acid sequence is from the subpopulation of maternal cells. In some embodiments, the nucleic acid sequence and / or the other nucleic acid sequence is a DNA sequence. In some embodiments, the nucleic acid sequence and / or the other nucleic acid sequence is a RNA sequence.

[0244] Non-limiting examples of sequencing platforms include Sanger sequencing, massively parallel sequencing (MPS), paired-end sequencing, high-throughput sequencing, next-generation sequencing (NGS), shotgun sequencing, single-molecule sequencing, nanopore sequencing, semiconductor sequencing, pyrosequencing, sequencing-by-synthesis (SBS), sequencing-by- ligation, sequencing-by-hybridization, and RNA-Seq (Illumina). In some embodiments, the sequencing is performed on a next generation sequencing platf orm. In some embodiments, the sequencing is performed on a massively parallel sequencing platform. In some embodiments, the sequencing is performed on a paired-end sequencing platform. In some embodiments, theWSGR Docket No. 62193-701.601 sequencing is performed on a polony sequencing platform. In some embodiments, the sequencing is performed on a high-throughput sequencing platform. In some embodiments, the sequencing is performed on a shotgun sequencing platform. In some embodiments, the sequencing is performed on a single-molecule sequencing platform. In some embodiments, the sequencing is performed on a nanopore sequencing platform. In some embodiments, the sequencing is performed on a semiconductor sequencing platform. In some embodiments, the sequencing is performed on a pyrosequencing platform. In some embodiments, the sequencing is performed on a sequencing-by-synthesis platform. In some embodiments, the sequencing is performed on a sequencing-by-ligation platform. In some embodiments, the sequencing is performed on a sequencing-by-hybridization platform. In some embodiments, the sequencing is performed on an RNA-SEQ (Illumina) platform. The sequencing can comprise nucleic acid amplification (e.g., of RNA or DNA molecules). In some embodiments, the entire genome is amplified prior to sequencing.

[0245] In some embodiments, the nucleic acid amplification is polymerase chain reaction (PCR). A suitable number of rounds of PCR (e.g., PCR, qPCR, reverse-transcriptase PCR, digital PCR, etc.) is performed to amplify an initial amount of nucleic acid (e.g., RNA or DNA) to a desired input quantity for subsequent sequencing. In some embodiments, the PCR is used for global amplification of target nucleic acids. This amplification can comprise using adapter sequences that are first ligated to different molecules followed by PCR amplification using universal primers. PCR can be performed using a kit of all the necessary tools. In some embodiments, only certain target nucleic acids within a population of nucleic acids can be amplified. Specific primers, for example, in conjunction with adapter ligation, can be used to amplify certain targets selectively for downstream sequencing. The PCR can comprise targeted amplification of one or more genomic loci, such as genomic loci associated with pregnancy - related states. The sequencing can comprise use of simultaneous reverse transcription (RT) and polymerase chain reaction (PCR), such as a OneStep RT-PCR kit protocol.

[0246] In some embodiments, the genetic analysis of fetal genetic material is PCR. In some embodiments, the PCT is quantitative PCR (qPCR). In some embodiments, the qPCR uses Taqman primer / probes sets. In some embodiments, the PCR is RT-PCR. In some embodiments, the PCR is droplet digital PCR (ddPCR). In some embodiments, the PCR is digital PCR. In some embodiments, the PCR analysis is a PCR-based mutation detection. In some embodiments, the PCR analysis is single nucleotide polymorphisms (SNP) genotyping arrays.

[0247] In some embodiments, the method further comprises collecting paternal cells from the father of the fetus. In some embodiments, the paternal cells from the father of the fetus areWSGR Docket No. 62193-701.601 collected from a biological sample taken from the father. In some examples, the biological sample is a blood sample. In some examples, the biological sample is a tissue and / or biopsy sample. In some examples, the biological sample is a biological fluid sample. In some examples, the biological fluid sample is blood, saliva, interstitial fluid, extracellular fluid, or any combination thereof.

[0248] In some embodiments, the paternal cells have from about one paternal cell type to about 10 paternal cell types. In some embodiments, the paternal cells have at least about one paternal cell types, at least about two paternal cell types, at least about three paternal cell types, at least about four paternal cell types, at least about five paternal cell types, at least about six paternal cell types, at least about seven paternal cell types, at least about eight paternal cell types, at least about nine paternal cell types, at least about ten paternal cell types, or more. In some embodiments, the paternal cells have at most about ten paternal cell types, at most about nine paternal cell types, at most about eight paternal cell types, at most about seven paternal cell types, at most about six paternal cell types, at most about five paternal cell types, at most about four paternal cell types, at most about three paternal cell types, at most abou t two paternal cell types, at most about one paternal cell type, or less. In some embodiments, the paternal cells have about one paternal cell types, about two paternal cell types, about three paternal cell types, about four paternal cell types, about five paternal cell types, about six paternal cell types, about seven paternal cell types, about eight paternal cell types, about nine paternal cell types, or about ten paternal cell types. Non-limiting examples of types of paternal cells include red blood cells, T cells, B cells, dendritic cells, macrophages, monocytes, erythrocytes, neutrophils, and lymphocytes.

[0249] In some embodiments, the paternal cells are isolated from the biological sample from the father. In some embodiments, the paternal cells are isolated from the biological sample from the father by removing non-nucleated cells (e.g., red blood cells), extracellular proteins, extracellular nucleic acid molecules, extracellular metabolites, or any combination thereof from the biological sample.

[0250] In some embodiments, the paternal cells can have one or more of the maternal biomarkers disclosed herein. In some embodiments, the paternal cells can couple to one or more of the isolation agents specific to maternal biomarkers disclosed herein.

[0251] In some embodiments, the paternal cells are quantified. In some embodiments, the paternal cells are quantified by flow cytometry. In some embodiments, the paternal cells are quantified by fluorescence-activated cell sorting (FACS) flow cytometry. In some embodiments, the paternal cells are isolated through magnetic-activated cell sorting (MACS). In someWSGR Docket No. 62193-701.601 embodiments, the paternal cells are quantified by microscopic microdissection. In some embodiments, the paternal cells are quantified by microscopic imaging. In some embodiments, the paternal cells are quantified by acoustic focusing flow cytometry. In some embodiments, the paternal cells are quantified by a hemocytometer. In some embodiments, the paternal cells are quantified by an automated cell counter. In some embodiments, the paternal cells are quantified by spectrophotometry.

[0252] In some embodiments, the paternal cells can be processed to generate sequencing data of the genome of the paternal cells. Processing the biological sample obtained from the subject can comprise (i) subjecting the paternal cells to conditions that are sufficient to isolate, enrich, or extract a plurality of fetal cell genetic material, and (ii) sequencing the plurality of paternal cell genetic material to generate a plurality of sequencing reads. The nucleic acid molecules can comprise ribonucleic acid (RNA) or deoxyribonucleic acid (DNA). The nucleic acid molecules can comprise DNA. The nucleic acid molecules can comprise RNA. The nucleic acid molecules (e.g., RNA or DNA) can be extracted from the paternal cells by a variety of methods, such as a DNeasy Blood and Tissue Kit, a PureLink Genomic DNA kit, or a Wizard® HMW DNA Extraction Kit. The extraction method can extract all RNA or DNA molecules from a sample.

[0253] In some embodiments, the genetic material from the paternal cells is genetically analyzed. In some embodiments, genetic material is analyzed via PCR, sequencing, hybridization methods, or any combination thereof. In some embodiments, the genetic material is analyzed via a whole genome scan. In some embodiments, the genetic material is analyzed via short tandem repeat analysis. In some embodiments, the genetic material is analyzed via microsatellite analysis.

[0254] In some embodiments, the genetic analysis is a hybridization method. In some embodiments, the hybridization method is a restriction fragment length polymorphism (RFLP), a southern blot analysis, a microarray, a fluorescence in situ hybridization (FISH), a comparative genomic hybridization, in situ hybridization, or any combination thereof.

[0255] In some embodiments, the genetic material from the paternal cells is amplified prior to genetic analysis. In some embodiments, the genetic material from the paternal cells is amplified to increase the amount of the genetic material from the paternal cells by between about 0% to about 500%. In some embodiments, the genetic material from the paternal cells is amplified to increase the amount of genetic material from the paternal cells by at least about 0%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 120%, at least about 140%, at least about 160%, at least about 180%, at least aboutWSGR Docket No. 62193-701.601 200%, at least about 250%, at least about 300%, at least about 350%, at least about 400%, at least about 450%, at least about 500%, or more. In some embodiments, the genetic material from the paternal cells is amplified to increase the amount of genetic material from the paternal cells by at most about 500%, at most about 450%, at most about 400%, at most about 350%, at most about 300%, at most about 250%, at most about 200%, at most about 180%, at most about 160%, at most about 140%, at most about 120%, at most about 100%, at most about 90%, at most about 80%, at most about 70%, at most about 60%, at most about 50%, at most about 40%, at most about 30%, at most about 20%, at most about 10%, at most about 0%, or less. In some embodiments, the genetic material from the paternal cells is amplified to increase the amount of genetic material from the paternal cells by about 0%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 120%, about 140%, about 160%, about 180%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, or about 500%.

[0256] In some embodiments, the fetal genetic material is analyzed in a genetic analysis. In some embodiments, the maternal genetic material is analyzed in a genetic analysis. In some embodiments, the fetal genetic material and the maternal genetic material are analyzed in a genetic analysis. In some embodiments, the paternal genetic material is analyzed in a genetic analysis. In some embodiments, the fetal genetic material, the maternal genetic material, and the paternal genetic material are analyzed in a genetic analysis.

[0257] In some embodiments, the genetic analysis is sequencing. In some embodiments, the method further comprises sequencing the nucleic acid molecules from the subpopulation of fetal cells. In some embodiments, the nucleic acid molecules from the subpopulation of fetal cells generate a nucleic acid sequence. In some embodiments, the nucleic acid sequence is from the subpopulation of fetal cells. In some embodiments, the method further comprises sequencing the nucleic acid molecules from the subpopulation of maternal cells. In some embodiments, the nucleic acid molecules from the subpopulation of maternal cells generate another nucleic acid sequence. In some embodiments, the other nucleic acid sequence is from the subpopulation of maternal cells. In some embodiments, the method further comprises sequencing the nucleic acid molecules from the paternal cells. In some embodiments, the nucleic acid molecules from the paternal cells generate a second nucleic acid sequence. In some embodiments, the second nucleic acid sequence is from the paternal cells. In some embodiments, the nucleic acid sequence, the second nucleic acid sequence, and / or the other nucleic acid sequence is a DNA sequence. In some embodiments, the nucleic acid sequence, the second nucleic acid sequence, and / or the other nucleic acid sequence is a RNA sequence.WSGR Docket No. 62193-701.601

[0258] Non-limiting examples of sequencing platforms include Sanger sequencing, massively parallel sequencing (MPS), paired-end sequencing, high-throughput sequencing, next-generation sequencing (NGS), shotgun sequencing, single-molecule sequencing, nanopore sequencing, semiconductor sequencing, pyrosequencing, sequencing-by-synthesis (SBS), sequencing-by- ligation, sequencing-by-hybridization, and RNA-Seq (Illumina). In some embodiments, the sequencing is performed on a next generation sequencing platform. In some embodiments, the sequencing is performed on a massively parallel sequencing platform. In some embodiments, the sequencing is performed on a paired-end sequencing platform. In some embodiments, the sequencing is performed on a polony sequencing platform. In some embodiments, the sequencing is performed on a high-throughput sequencing platform. In some embodiments, the sequencing is performed on a shotgun sequencing platform. In some embodiments, the sequencing is performed on a single-molecule sequencing platform. In some embodiments, the sequencing is performed on a nanopore sequencing platform. In some embodiments, the sequencing is performed on a semiconductor sequencing platform. In some embodiments, the sequencing is performed on a pyrosequencing platform. In some embodiments, the sequencing is performed on a sequencing-by-synthesis platform. In some embodiments, the sequencing is performed on a sequencing-by-ligation platform. In some embodiments, the sequencing is performed on a sequencing-by-hybridization platform. In some embodiments, the sequencing is performed on an RNA-SEQ (Illumina) platform. The sequencing can comprise nucleic acid amplification (e.g., of RNA or DNA molecules). In some embodiments, the entire genome is amplified prior to sequencing.

[0259] In some embodiments, the nucleic acid amplification is polymerase chain reaction (PCR). A suitable number of rounds of PCR (e.g., PCR, qPCR, reverse-transcriptase PCR, digital PCR, etc.) is performed to amplify an initial amount of nucleic acid (e.g., RNA or DNA) to a desired input quantity for subsequent sequencing. In some embodiments, the PCR is used for global amplification of target nucleic acids. This amplification can comprise using adapter sequences that are first ligated to different molecules followed by PCR amplification using universal primers. PCR can be performed using a kit of all the necessary tools. In some embodiments, only certain target nucleic acids within a population of nucleic acids can be amplified. Specific primers, for example, in conjunction with adapter ligation, can be used to amplify certain targets selectively for downstream sequencing. The PCR can comprise targeted amplification of one or more genomic loci, such as genomic loci associated with pregnancy - related states. The sequencing can comprise use of simultaneous reverse transcription (RT) and polymerase chain reaction (PCR), such as a OneStep RT-PCR kit protocol.WSGR Docket No. 62193-701.601

[0260] In some embodiments, the genetic analysis of fetal genetic material, the maternal genetic material, and / or the paternal genetic material is PCR. In some embodiments, the PCT is quantitative PCR (qPCR). In some embodiments, the qPCR uses Taqman primer / probes sets. In some embodiments, the PCR is RT-PCR. In some embodiments, the PCR is droplet digital PCR (ddPCR). In some embodiments, the PCR is digital PCR. In some embodiments, the PCR analysis is a PCR-based mutation detection. In some embodiments, the PCR analysis is single nucleotide polymorphisms (SNP) genotyping arrays.

[0261] After subjecting the nucleic acid molecules to sequencing, suitable bioinformatics processes can be performed on the sequence reads to generate the data indicative of the presence or absence of genetic disorders in the fetus (or fetuses). In some embodiments, the sequence reads are aligned to one or more reference genomes (e.g., a genome of one or more species such as a human genome). The aligned sequence reads can be quantified at one or more genomic loci to identify the presence or absence of the genetic disorder.

[0262] In some embodiments, the subpopulation of maternal cells is quantified. In some embodiments, the subpopulation of maternal cells is quantified by flow cytometry. In some embodiments, the subpopulation of maternal cells is quantified by fluorescence-activated cell sorting (FACS) flow cytometry. In some embodiments, the ex vivo population of cells is isolated through magnetic-activated cell sorting (MACS). In some embodiments, the subpopulation of maternal cells is quantified by microscopic microdissection. In some embodiments, the subpopulation of maternal cells is quantified by microscopic imaging. In some embodiments, the subpopulation of maternal cells is quantified by acoustic focusing flow cytometry. In some embodiments, the subpopulation of maternal cells is quantified by a hemocytometer. In some embodiments, the subpopulation of maternal cells is quantified by an automated cell counter. In some embodiments, the subpopulation of maternal cells is quantified by spectrophotometry.

[0263] In some embodiments, the subpopulation of fetal cells is quantified. In some embodiments, the subpopulation of fetal cells is quantified by flow cytometry. In some embodiments, the subpopulation of fetal cells is quantified by fluorescence-activated cell sorting (FACS) flow cytometry. In some embodiments, the ex vivo population of cells is isolated through magnetic-activated cell sorting (MACS). In some embodiments, the subpopulation of fetal cells is quantified by microscopic microdissection. In some embodiments, the subpopulation of fetal cells is quantified by microscopic imaging. In some embodiments, the subpopulation of fetal cells is quantified by acoustic focusing flow cytometry. In some embodiments, the subpopulation of fetal cells is quantified by a hemocytometer. In some embodiments, the subpopulation of fetal cells is quantified by an automated cell counter. InWSGR Docket No. 62193-701.601 some embodiments, the subpopulation of fetal cells is quantified by spectrophotometry. In some embodiments, the subpopulation of maternal cells and the subpopulation of fetal cells are quantified. In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells are quantified by flow cytometry.

[0264] In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells are quantified by fluorescence-activated cell sorting (FACS) flow cytometry. In some embodiments, the ex vivo population of cells is isolated through magnetic-activated cell sorting (MACS). In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells are quantified by microscopic microdissection. In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells are quantified by microscopic imaging. In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells are quantified by acoustic focusing flow cytometry. In some embodiments, subpopulation of fetal cells and the subpopulation of maternal cells are quantified by a hemocytometer. In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells are quantified by an automated cell counter. In some embodiments, the subpopulation of fetal cells and the subpopulation of maternal cells are quantified by spectrophotometry.

[0265] In some embodiments, the present disclosure provides a method of performing an assay on an ex vivo population of cells. In some embodiments, the ex vivo population of cells is isolated from a subject. In some embodiments, the ex vivo population of cells are isolated from a biological sample taken from the subject. In some embodiments, the biological sample is a blood sample. In some embodiments, the blood sample is plasma. In some embodiments, the blood sample is serum. In some embodiments, the blood sample is whole blood. In some embodiments, the blood sample is whole blood depleted of red blood cells. In some embodiments, the blood sample is a buffy coat. In some embodiments, the blood sample is peripheral blood mononuclear cells. In some embodiments, the blood sample is nucleated white blood cells. In some embodiments, the biological sample is a tissue and / or biopsy sample. In some embodiments, the tissue and / or biopsy sample is from the uterus, placenta, umbilical cord, or any combination thereof. In some embodiments, the biological sample is a biological fluid sample. In some embodiments, the biological fluid sample is blood, saliva, interstitial fluid, umbilical cord blood, extracellular fluid, or any combination thereof .

[0266] In some embodiments, the sample is from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a mouse. In some embodiments, the subject is a rat. In some embodiments, the subject is a non-human primate. In some embodiments, theWSGR Docket No. 62193-701.601 subject is a human. In some embodiments, the sample is from a pregnant subject. In some embodiments, the sample from the pregnant subject is a maternal biological sample. In some embodiments, the ex vivo population of cells has a subpopulation of fetal cells. In some embodiments, the ex vivo population of cell has a subpopulation of maternal cells. In some embodiments, the ex vivo population of cells has a subpopulation of fetal cells and a subpopulation of maternal cells.

[0267] In some embodiments, the subpopulation of fetal cells is from about 50% to about 100% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is from about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at most about 100%, at most about 95%, at most about 90%, at most about 85%, at most about 80%, at most about 75%, at most about 70%, at most about 65%, at most about 60%, at most about 55%, at most about 50%, or less of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 50% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 60% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 70% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 80% of cells in the ex vivo population of cells. In some embodiments, the subpopulation of fetal cells is at least about 90% of cells in the ex vivo population of cells.

[0268] In some embodiments, the method further comprising depleting red blood cells from the ex vivo population of cells.

[0269] In some embodiments, the assay is a cell morphology assay. Non-limiting examples of cell morphology assays include immunofluorescence, scanning electron microscopy, and hematoxylin-Eosin (HE) staining. In some embodiments, the cell morphology assay can determine a size of a cell, a shape of a cell, presence or absence of organelles in a cell, presence or absence of cell membrane of a cell, number of organelles in a cell, or any combination thereof. In some embodiments, the assay is a cell counting assay. Non-limiting examples of cell counting assays include flow cytometry, FACS, MACS, acoustic focusing flow cytometry, hemocytometry, microscopic microdissection, microscopic imaging, and spectrophotometry. In some embodiments, the assay is a cell viability assay. Non-limiting examples of cell viabilityWSGR Docket No. 62193-701.601 assays include cell metabolism assays, ATP detection assays, tetrazolium reduction assays, and protease activity assays. In some embodiments, the assay is a cell proliferation assay. Non - limiting examples of cell proliferation assays include DNA synthesis proliferation assays, generational analysis by dye dilution assays, and DNA content quantitation cell proliferation assays. In some embodiments, the assay is a cytotoxicity assay. Non-limiting examples of cytotoxicity assays include lactate dehydrogenase assays, DNA-binding dye assays, and Annexin V staining.

[0270] In some embodiments, the assay indicates a likelihood that the ex vivo population of cells is susceptible to a disease or condition. In some embodiments, the assay indicates a likelihood that the subpopulation of maternal cells is susceptible to a disease. In some embodiments, the assay indicates a likelihood that the subpopulation of fetal cells is susceptible to a disease. In some embodiments, the assay indicates that a fetus associated with subpopulation of fetal cells is susceptible to a disease. In some embodiments, the disease is a genetic condition.

[0271] In some embodiments, the results of the assay is compared to information from one or more databases. In some embodiments, the one or more databases include information related to fetal disorders, genetic disorders, genetic conditions, fetal or any combination thereof. Non - limiting examples of the information from the one or more databases include genetic sequencing information from fetal cells affected by a genetic disorder or condition, morphological characteristics of fetal cells affected by a genetic disorder or condition, chromosomal make-up of fetal cells affected by a genetic disorder or condition, gene expression information from the subpopulation of fetal cells affected by a genetic disorder or condition, protein expression levels from fetal cells affected by a genetic disorder or condition, functional characteristics of fetal cells affected by a genetic disorder or condition, or any combination thereof. In some embodiments, the one or more databases are preexisting databases. In some embodiments, the one or more databases are databases currently being generated. In some embodiments, the results of the assay are compared to the one or more databases to determine if the fetus has a genetic disorder or a genetic condition. In some embodiments, the comparison of the results of the assay and the one or more databases can be provided to the mother. In some embodiments, the comparison of the results of the assay and the one or more databases can be provided to a clinician. In some embodiments, the comparison of the results of the assay and the one or more databases can be provided to the mother and a clinician. In some embodiments, the comparison of the results of the assay and the one or more databases generates a diagnosis of the one or more genetic conditions or disorders. In some embodiments, if the results from the assay is the same as the information from the one or more databases, then the comparison generates aWSGR Docket No. 62193-701.601 diagnosis of one or more genetic conditions or disorders. In some embodiments, if the results from the assay is different from the information from the one or more databases, the then comparison may not generate a diagnosis of one or more genetic conditions or disorders.

[0272] Non-limiting examples of genetic conditions or disorders include 1p36 deletion syndrome, 1q21.1 deletion syndrome, 2q37 deletion syndrome, 5q deletion, 5, 10 - methenyltetrahydrofolate synthestase deficiency, 17q12 microdeletion syndrome, 17q12 microduplication syndrome, 18p deletion syndrome, 21-hydroxylase deficiency, alpha 1- antitrypsin deficiency, achalasia-addisonianism-alacrima (AAA) syndrome, Aarskog-Scott syndrome, ABCD syndrome, Absnece deformity of leg-cataract, Aceruloplasminemia, Acheiropodia, Achondrogenesis type II, achondroplasia, acute intermittent porphyria, adenylosuccinate lyase deficiency, adrenoleukodyst97estatiolagille syndrome, ADULT syndrome, Aicardi-Goutieres syndrome, Albinism, Alexander disease, Alfi’s syndrome, alkaptonuria, Alport syndrome, alternating hemiplegia of childhood, aortic arch anomaly, peculiar facies, Amish lethal microcephaly, amyotrophic lateral sclerosis, angel-shaped phalango-epiphyseal dysplasia, Alstrom syndrome, Alzheimer’s syndrome, amelogenesis imperfecta, aminolevulinic acid dehydratase deficiency porphyria, androgen insensitivity syndrome, angelman syndrome, aphalangy-syndactyly-microcephaly syndrome, apert syndrome, arthrogryposis, ataxia telangiectasia, axenfeld syndrome, Bainbridge-ropers syndrome, breae- stevenson cutis gyrate syndrome, beckwith-wiedermann syndrome, Benjamin syndrome, biotinidase deficiency, Bjornstad syndrome, Blepharophimosis syndromes, Bloom syndrome, Birt-Hogg-Dube syndrome, Brody myopathy, Brunner syndrome, CADASIL syndrome, Cat eye syndrome, CRASIL syndrome, chronic granulomatous disorder, campomelic dysplasia, captodactyly-taurinuria syndrome, Canavan disease, Carpenter syndrome, CDKL5 deficiency disorder, cerebral dysgenesis-neuropathy-ichthyosis-keratoderma syndrome (CEDNIK), cleft palate short stature vertebral anomalies syndrome, congenital muscular dystrophy -infantile cataract-hypogonadism syndrome, cystic fibrosis, Charcot-Marie-Tooth disease, CHARGE syndrome, Chediak-Higashi syndrome, Grebe-type chonodrodysplasia, cleidocranial dysostosis, Cockayne syndrome, Coffin-Lowry syndrome, Cohen syndrome, type II collagenopathy, type XI collagenopathy, congenital insensitivity to pain with anhidrosis (CIPA), congenital muscular dystrophy, cornelia de Lange syndrome, Cowden syndrome, CPO deficiency, Cranio-lenticulo- sutural dysplasia, Cri du chat, Crohn’s disease, Crouzon syndrome, Crouzonodermoskeletal syndrome, Currarino syndrome, Darier’s disease, Dent’s disease, Denys-Drash syndrome, De Grouchy syndrome, dolichonychia, Down Syndrome, DiGeorge Syndrome, Distal hereditary motor neuropathies, distal muscular dystrophy, Duchenne muscular dystrophy, DravetWSGR Docket No. 62193-701.601 syndrome, Ectrodactyly-polydactyly syndrome, Edwards Syndrome, Ehlers-Danlos syndrome, Emanuel syndrome, Emery-Dreifuss syndrome, Epidermolysis bullosa, erythropoietic protoporphyria, Fanconi amenia, Fabry disease, Factor V Leiden thrombophilia, fatal familial insomnia, familial adenomatous polyposis, familial dysautonomia, Familial Creutzfeld -Jakob disease, familial episodic pain syndrome, familial thoracic aortic aneurysm and aortic dissection, Feingold syndrome, FG syndrome, FBXW7 neurodevelopmental syndrome, Fibular aplasia - ectrodactyly syndrome, Fine-Lubinsky syndrome, Fragile X syndrome, Friedreich’s ataxia, G6PD deficiency, glactosemia, Gaucher disease, Gerstmann-Straussler-Scheinker syndrome, Gillespie syndrome, type I glutaric aciduria, type II glutaric aciduria, GRACILE syndrome, GRIN2B-related neurodevelopmental disorder, Griscelli syndrome, Gustavson syndrome, Hailey-Hailey disease, Harlequin type ichthyosis, Hemochromatosis type 1, Hemochromatosis type 2A, Hemochromatosis type 2B, Hemochromatosis type 3, Hemochromatosis 4, Hemochromatosis 5, hemophilia, hepatoerythropoietic porphyria, hereditary coproporphyria, hereditary hemorrhagic telangiectasia, hereditary inclusion body myopathy, hereditary multiple exostoses, hereditary spastic paraplegia, Hermansky-Pudiak syndrome, hereditary neuropathy with liability to pressure palsies (HNPP), heterotaxy, homocystinuria, Huntington’s disease, Hunter syndrome, Hurler syndrome, Hutchinson-Gilford progeria syndrome, hyperlysinemia, primary hyperoxaluria, hyperphenylalainemia, Tangier disease, hypochondrogenesis, hypochondroplasia, immunodeficiency-centrometric instability-facial anomalies syndrome (ICF), incontinentia pigmenti, infantile cerebral and cerebellar atrophy with postnatal progressive microcephaly, ischiopatellar dysplasia, isodicentric 15, PRICKLE1-related progressive myoclonus epilepsy with ataxia, Jackson-Weiss syndrome, Jacobsen syndrome, Joubert syndrome, Juvenile-onset dystonia, Juvenile primary lateral sclerosis, Keloid disorder, KIF1A-associated neurological disorder, Kleefstra syndrome, Kniest dysplasia, Kosaki overgrowth syndrome, Krabbe disease, Kufor-Rakeb syndrome, LCAT deficiency, Lesch-Nyhan syndrome, Li-Fraumeni syndrome, Limb-Gridle Muscular Dystrophy, Lynch syndrome, lipoprotein lipase deficiency, malignany hyperthermia, Maple syrup urine disease, Marfan syndrome, Maroteaux-Lamy syndrome, McCune-Albright syndrome, McLeod syndrome, MEDNIK syndrome, familial Mediterranean fever, Mekes disease, methemoglobinemia, methylmalonic acidemia, Micro syndrome, microcephaly, Miller-Dieker syndrome, Morquio syndrome, Mowat-Wilson syndrome, Muenke syndrome, type 1 multiple endocrine neoplasia, type 2 multiple endocrine neoplasia, muscular dystrophy, Becker muscular dystrophy, myostatin-related muscle hypertrophy, myotonic dystrophy, Natowicz syndrome, Nedamss neurodevelopment disorder, type I neurofibromatosis, type II neurofibromatosis, Niemann-PickWSGR Docket No. 62193-701.601 disease, Nonketotic hyperglycinemia, nonsyndromic deafness, noonan syndrome, Norman - Roberts syndrome, Ogden syndrome, Omenn syndrome, Osteogenesis imperfecta, Ostravik - Lindemann-Solberg syndrome, Pantothenate kinase-associated neurodegeneration, Patau syndrome, PCC deficiency, porphyria cutanea tarda, Pendred syndrome, Peutz -Jeghers syndrome, Pfeiffer syndrome, Phelan-McDermid syndrome, Phenylketouria, Pipecolic acidemia, Pitt-Hopkins syndrome, Polycystic kidney disease, polycystic ovary syndrome, porphyria, Prader-Willi syndrome, Primary ciliary dyskinesia, primary pulmonary hypertension, protein C deficiency, protein S deficiency, proximal 18q deletion syndrome, Pseudo-Gaucher disease, pseudoxanthoma elasticum, Retinitis pigmentosa, Rett syndrome, Roberts syndrome, Rubinstein-Taybi syndrome, Sandhoff disease, Sanfilippo syndrome, Schwartz-Jampel syndrome, Sjogren-Larsson syndrome, Skin fragility-wooly hair-palmoplantar keratoderma syndrome, Spondyloepiphyseal dysplasia congenita, Shprintzen-Goldbery syndrome, Sickle cell anemia, Siderius X-linked mental retardation syndrome, Sideroblastic anemia, Sly syndrome, Smith-Lemli-Optiz syndrome, Smith-Magenis syndrome, Synder-Robinson syndrome, Spinal muscular atrophy, Spinocerebellar ataxia, split hand split foot-nystagmus syndrome, SSB syndrome, Stargardt disease, Stickler syndrome, Strudwick syndrome, Tay-Sachs disease, tetrahydrobiopterin deficiency, thanatophoric dysplasia, thickened earlobes-conductive deafness syndrome, Treacher Collins syndrome, Tuberous sclerosis complex, Turner syndrome, Usher syndrome, Variegate porphyria, Vijoen-Kallis-Voges syndrome, von Hippel-Lindau disease, von Willebrand disease, Waardenburg syndrome, Warkany syndrome 2, Weissenbacher-Zweymüller syndrome, Weyer’s ulnar ray / oligodactyly syndrome, Williams syndrome, Wilson disease, Woodhouse-Sakati syndrome, Wolf-Hirschhorn syndrome, Xeroderma pigmentosum, Fragile X syndrome, X-linked spinal-bulbar muscle atrophy, Xp11.2 duplication syndrome, X-linked severe combined immunodeficiency, X-linked sideroblastic anemia, triple X syndrome, XXXX syndrome, XXXXX syndrome, XXXXY syndrome, XYY syndrome, XXYY syndrome, XYYY syndrome, XXXY syndrome, XYYYY syndrome, and Zellweger syndrome.

[0273] In some embodiments, the genetic condition is down syndrome. In some embodiments, the genetic disorder is Fragile X syndrome. In some embodiments, the genetic disorder is triple X syndrome. In some embodiments, the genetic disorder is XXXX syndrome. In some embodiments, the genetic disorder is XXXXX syndrome. In some embodiments, the genetic disorder is XXXXY. In some embodiments, the genetic disorder is XYY syndrome. In some embodiments, the genetic disorder is XYY syndrome. In some embodiments, the genetic disorder is XXYY syndrome. In some embodiments, the genetic disorder is XYYY. In some embodiments, the genetic disorder is XXXY syndrome. In some embodiments, the geneticWSGR Docket No. 62193-701.601 disorder is XYYYY. In some embodiments, the genetic disorder is Tay-Sachs disease. In some embodiments, the genetic disorder is Angelman syndrome. In some embodiments, the genetic disorder is Cystic fibrosis. In some embodiments, the genetic disorder is DiGeorge syndrome. In some embodiments, the genetic disorder is muscular dystrophy. In some embodiments, the genetic disorder is Duchenne muscular dystrophy. In some embodiments, the genetic disorder is fetal familial insomnia. In some embodiments, the genetic disorder is Creutzfeld-Jakob disease. In some embodiments, the genetic disorder is Gerstmann-Sträussler-Scheinker syndrome. In some embodiments, the genetic disorder is Hailey-Hailey disease. In some embodiments, the genetic disorder is Hemophilia. In some embodiments, the genetic disorder is Huntington’s disease. In some embodiments, the genetic disorder is Marfan syndrome. In some embodiments, the genetic disorder is Prader-Willi syndrome. In some embodiments, the genetic disorder is Sickle Cell disease. In some embodiments, the genetic disorder is Turner syndrome. In some embodiments, the genetic disorder is von Hippel-Lindau disease.

[0274] In some embodiments, the genetic condition is a chromosomal abnormality. Non- limiting examples of chromosomal abnormalities include aneuploidy, translocation, unbalanced translocation, rearrangement, subtelomeric rearrangement, unbalance chromosomal rearrangement, unbalance subtelomeric rearrangement, deletion, inversions, unbalanced inversions, duplication, telomere instability, telomere shortening, single nucleotide substitution, micro deletion, micro-insertion, short deletions, short insertion, multi-nucleotide changes, DNA methylation and, and loss of imprint (LOI). Non-limiting examples of chromosomal aneuploidy include trisomy 21, trisomy 18, trisomy 13, trisomy 16, polysomies, monosomies, and XXX. In some embodiments, the chromosomal aneuploidy is a complete monosomy. In some embodiments, the complete monosomy is monosomy X, monosomy 21, monosomy 22, monosomy 16, or monosomy 15. In some embodiments, the chromosomal aneuploidy is a partial monosomy. In some embodiments, the partial monosomy is monosomy X, monosomy 21, monosomy 22, monosomy 16, or monosomy 15. In some embodiments, the chromosomal abnormality is trisomy 18. In some embodiments, the chromosomal abnormality is trisomy 13.

[0275] In some embodiments, the subpopulation of fetal cells are used to perform a fetal genetic risk analysis. In some embodiments, the fetal genetic risk analysis determines whether an adverse health factor to the pregnancy exists. In some embodiments, the fetal genetic risk analysis determines whether an absence of an adverse health factor to the pregnancy exists. In some embodiments, the fetal genetic risk analysis determines a presence of of a genetic disorder or condition in the fetus. In some embodiments, the fetal genetic risk analysis determines an absence of a genetic disorder or condition in the fetus. In some embodiments, the fetal geneticWSGR Docket No. 62193-701.601 risk analysis determines the current health of the fetus. In some examples, the fetal genetic risk analysis determines whether the fetus has a low risk, a medium risk, or a high risk of developing health complications during the pregnancy. In some embodiments, a low risk of developing health complications during the pregnancy can mean an unlikelihood (e.g., less than about 30% probability) that the fetus develops health complications during the pregnancy. In some embodiments, a medium risk of developing health complications during the pregnancy can mean a possibility (e.g., between about 30% to about 60% probability) that the fetus develops health complications during the pregnancy. In some embodiments, a high risk of developing health complications during the pregnancy can mean a high likelihood (e.g., greater than about 60% probability) that the fetus develops health complications during the pregnancy. In some embodiments, the fetal genetic risk analysis determines the current health of the mother during the pregnancy. In some embodiments, the fetal genetic risk analysis determines whether the mother has a low risk, a medium risk, or a high risk of developing health complications during the pregnancy. In some embodiments, a low risk of developing health complications during the pregnancy can mean an unlikelihood (e.g., less than about 30% probability) that the mother develops health complications during the pregnancy. In some embodiments, a medium risk of developing health complications during the pregnancy can mean that a possibility (e.g., between about 30% to about 60% probability) that the mother develops health complications during the pregnancy. In some embodiments, a high risk of developing health complications during the pregnancy can mean a high likelihood (e.g., greater than about 60% probability) that the mother develops health complications during the pregnancy. In some embodiments, the fetal genetic risk analysis determines the current risk level of a miscarriage in the mother. In some embodiments, the fetal genetic risk analysis is performed via performing one or more of the assays disclosed herein. In some embodiments, the fetal genetic risk analysis performed via a sequencing assay on the subpopulation of the fetal cells.

[0276] In some embodiments, the one or more assays performed can be a genetic assay. In some examples, the genetic assay can be PCR, whole genome sequencing (WGS), targeted genomic sequencing, targeted ribonucleic acid sequencing, hybridization methods, short tandem repeat analysis, microsatellite analysis, or any combination thereof. In some embodiments, the one or more assays performed can be a cell morphology assay. In some embodiments, the one or more assays can be a cell counting assay. In some embodiments, the one or more assays can be a cell viability assay. In some embodiments, the one or more assays can be a cell proliferation assay. In some embodiments, the one or more assays can be a cytotoxicity assay. In some embodiments, the one or more assays can be a cell function assay. In some examples, the cellWSGR Docket No. 62193-701.601 function assay can include a cell migration assay, a cell invasion assay, an angiogenesis a...

Claims

WSGR Docket No. 62193-701.601 CLAIMS WHAT IS CLAIMED IS:

1. A composition comprising: an ex vivo population of cells from a maternal blood sample, wherein the ex vivo population of cells comprises a subpopulation of maternal cells and a subpopulation of fetal cells, wherein the subpopulation of fetal cells is at least about 75% of cells in the ex vivo population of cells.

2. The composition of claim 1, wherein the subpopulation of fetal cells is at least about 80% of the cells in the ex vivo population of cells.

3. The composition of claim 1 or claim 2, wherein the subpopulation of fetal cells is at least about 90% of the cells in the ex vivo population of cells.

4. The composition of any one of claims 1-3, wherein the subpopulation of maternal cells is no greater than about 30% of the cells in the ex vivo population of cells.

5. The composition of any one of claims 1-4, wherein the subpopulation of maternal cells is no greater than about 20% of the cells in the ex vivo population of cells.

6. The composition of any one of claims 1-5, wherein the subpopulation of maternal cells is no greater than about 10% of the cells in the ex vivo population of cells.

7. The composition of any one of claims 1-6, wherein the subpopulation of fetal cells comprises trophoblasts.

8. The composition of any one of claims 1-7, further comprising a second subpopulation of fetal cells, wherein the second subpopulation of fetal cells is different in cell type from the subpopulation of fetal cells.

9. The composition of claim 8, wherein the second subpopulation of fetal cells comprises erythroblasts.

10. The composition of claim 8, wherein the second subpopulation of fetal cells comprises megakaryocytes.

11. The composition of claim 8, wherein the second subpopulation of fetal cells comprises vascular endothelial cells.WSGR Docket No. 62193-701.601 12. The composition of claim 8, wherein the second subpopulation of fetal cells comprises fetal stromal cells.

13. The composition of claim 8, wherein the subpopulation of fetal cells comprises early gestation fetal cells, and the second subpopulation of fetal cells comprises later gestation fetal cells.

14. The composition of any one of claims 1-13, wherein the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by flow cytometry.

15. A method comprising: sequencing a nucleic acid sequence, wherein the nucleic acid sequence is from an ex vivo population of cells from a maternal blood sample, wherein the ex vivo population of cells comprises a subpopulation of maternal cells and a subpopulation of fetal cells, wherein the subpopulation of fetal cells is at least about 70% of cells in the ex vivo population of cells.

16. The method of claim 15, wherein the subpopulation of fetal cells is at least about 80% of the cells in the ex vivo population of cells.

17. The method of claim 15 or claim 16, wherein the subpopulation of fetal cells is at least about 90% of the cells in the ex vivo population of cells.

18. The method of any one of claims 15-17, wherein the subpopulation of maternal cells is no greater than about 30% of the cells in the ex vivo population of cells.

19. The method of any one of claims 15-18, wherein the subpopulation of maternal cells is no greater than about 20% of the cells in the ex vivo population of cells.

20. The method of any one of claims 15-19, wherein the subpopulation of maternal cells is no greater than about 10% of the cells in the ex vivo population of cells.

21. The method of any one of claims 15-20, further comprising, prior to the sequencing the nucleic acid sequence, extracting the subpopulation of maternal cells and the subpopulatio n of fetal cells from the maternal blood sample to provide an extract, wherein the extract comprises the subpopulation of maternal cells and the subpopulation of fetal cells from the maternal blood sample.WSGR Docket No. 62193-701.601 22. The method of claim 21, further comprising enriching the subpopulation of fetal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

23. The method of claim 21, further comprising sorting the subpopulation of fetal cells in the extract from the subpopulation of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

24. The method of any one of claims 15-23, wherein prior to the sequencing the nucleic acid sequence, the subpopulation of maternal cells and the subpopulation of fetal cells are extracted from the maternal blood sample to provide an extract, wherein the extract comprises the subpopulation of maternal cells and the subpopulation of fetal cells from the maternal blood sample.

25. The method of claim 24, wherein the subpopulation of fetal cells in the extract is enriched from the subpopulation of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

26. The method of claim 24, wherein the subpopulation of fetal cells in the extract is sorted from the subpopulation of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

27. The method of any one of claims 15-26, wherein the subpopulation of fetal cells comprises trophoblasts.

28. The method of any one of claims 15-27, wherein the ex vivo population of cells further comprises a second subpopulation of fetal cells, wherein the second subpopulation of fetal cells is different in cell type from the first subpopulation of fetal cells.

29. The method of claim 28, wherein the second subpopulation of fetal cells comprises erythroblasts.

30. The method of claim 28, wherein the second subpopulation of fetal cells comprises megakaryocytes.

31. The method of claim 28, wherein the second subpopulation of fetal cells comprises vascular endothelial cells.WSGR Docket No. 62193-701.601 32. The method of claim 28, wherein the second subpopulation of fetal cells comprises stromal cells.

33. The method of claim 28, wherein the subpopulation of fetal cells comprises early gestation fetal cells and the second subpopulation of fetal cells comprises later gestation fetal cells.

34. The method of any one of claims 15-33, further comprising, prior to the sequencing the nucleic acid sequence, isolating the subpopulation of fetal cells from the maternal blood sample by contacting the maternal blood sample with a first isolation agent and coupling the first isolation agent to a first biomarker in the subpopulation of fetal cells.

35. The method of claim 34, further comprising incubating the maternal blood sample after the contacting.

36. The method of claim 35, wherein the first isolation agent is a nucleic acid probe.

37. The method of claim 35, wherein the first isolation agent is a ligand .

38. The method of claim 35, wherein the first isolation agent couples to an intracellular biomarker.

39. The method of claim 35, wherein the first isolation agent couples to a cell surface biomarker.

40. The method of claim 35, wherein the first isolation agent couples to a soluble cell surface biomarker.

41. The method of claim 35, wherein the first isolation agent couples to a nucleic acid biomarker.

42. The method of any one of claims 15-41, wherein prior to the sequencing the nucleic acid sequence, the subpopulation of fetal cells is isolated from the maternal blood sample, wherein the maternal blood sample is contacted with a first isolation agent and the first isolation agent couples to a first biomarker in the subpopulation of fetal cells.

43. The method of claim 42, wherein after the contacting, the maternal blood sample is incubated.

44. The method of claim 42, wherein the first isolation agent is a nucleic acid probe.WSGR Docket No. 62193-701.601 45. The method of claim 42, wherein the first isolation agent is a ligand.

46. The method of claim 42, wherein the first isolation agent couples to a cell surface biomarker.

47. The method of claim 42, wherein the first isolation agent couples to a soluble cell surface biomarker.

48. The method of claim 42, wherein the first isolation agent couples to an intracellular biomarker.

49. The method of claim 42, wherein the first isolation agent couples to a nucleic acid biomarker.

50. The method of any one of claims 15-49, wherein prior to the sequencing the nucleic acid sequence, the subpopulation of maternal cells is isolated from the maternal blood sample, wherein the maternal blood sample is contacted with a second isolation agent and the second isolation agent couples to a second biomarker in the subpopulation of maternal cells.

51. The method of claim 50, wherein after the contacting, the maternal blood sample is incubated.

52. The method of claim 50, wherein the second isolation agent is a nucleic acid probe.

53. The method of claim 50, wherein the second isolation agent is a ligand.

54. The method of claim 50, wherein the second isolation agent couples to a cell surface biomarker.

55. The method of claim 50, wherein the second isolation agent couples to a soluble cell surface biomarker.

56. The method of claim 50, wherein the second isolation agent couples to an intracellular biomarker.

57. The method of claim 50, wherein the second isolation agent couples to a nucleic acid biomarker.

58. The method of any one of claims 15-57, further comprising, prior to the sequencing the nucleic acid sequence, isolating the subpopulation of maternal cells from the maternal bloodWSGR Docket No. 62193-701.601 sample by contacting the maternal blood sample with a second isolation agent and coupling the second isolation agent to a second biomarker in the subpopulation of maternal cells.

59. The method of claim 58, further comprising incubating the maternal blood sample after the contacting.

60. The method of claim 58, wherein the second isolation agent is a nucleic acid probe.

61. The method of claim 58, wherein the second isolation agent is a ligand.

62. The method of claim 58, wherein the second isolation agent couples to a cell surface biomarker.

63. The method of claim 58, wherein the second isolation agent couples to a soluble cell surface biomarker.

64. The method of claim 58, wherein the second isolation agent couples to an intracellular biomarker.

65. The method of claim 58, wherein the second isolation agent couples to a nucleic acid biomarker.

66. The method of any one of claims 15-65, wherein the sequencing is performed on a next generation sequencing platform.

67. The method of any one of claims 15-66, wherein the nucleic acid sequence is from the subpopulation of fetal cells.

68. The method of claim 67, wherein the nucleic acid sequence is a DNA sequence.

69. The method of claim 67, wherein the nucleic acid sequence is a RNA sequence.

70. The method of any one of claims 15-69, wherein the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by flow cytometry.

71. A method comprising: performing an assay on an ex vivo population of cells from a maternal blood sample, wherein the ex vivo population of cells comprises a subpopulation of maternal cells and a subpopulation of fetal cells, wherein the subpopulation of fetal cells is at least about 70% of cells in the ex vivo population of cells.WSGR Docket No. 62193-701.601 72. The method of claim 71, wherein the assay is a cell morphology assay.

73. The method of claim 71, wherein the assay is a cell counting assay.

74. The method of claim 71, wherein the assay is a cell viability assay.

75. The method of claim 71, wherein the assay is a cell proliferation assay.

76. The method of claim 71, wherein the assay is a cytotoxicity assay.

77. The method of claim 71, wherein the assay indicates a likelihood that a fetus associated with the subpopulation of fetal cells is susceptible to a genetic condition.

78. The method of claim 77, wherein the genetic condition is down syndrome.

79. The method of claim 77, wherein the genetic condition is cystic fibrosis.

80. The method of claim 77, wherein the genetic condition is muscular dystrophy.

81. The method of claim 77, wherein the genetic condition is sickle cell disease.

82. The method of claim 77, wherein the genetic condition is Tay-Sachs disease.

83. The method of any one of claims 71-82, wherein the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by flow cytometry.

84. The method of any one of claims 71-83, wherein the subpopulation of maternal cells and the subpopulation of fetal cells in the ex vivo population of cells is quantified by microscopy.

85. A method comprising: (a) sequencing a first nucleic acid sequence obtained from a first type of fetal cell from an ex vivo population of fetal cells; (b) sequencing a second nucleic acid sequence obtained from a second type of fetal cell from the ex vivo population of fetal cells; (c) sequencing a third nucleic acid sequence obtained from a third type of fetal cell from the ex vivo population of fetal cells; (d) sequencing a fourth nucleic acid sequence obtained from a fourth type of fetal cell from the ex vivo population of fetal cells;WSGR Docket No. 62193-701.601 (e) sequencing a fifth nucleic acid sequence obtained from a fifth type of fetal cell from the ex vivo population of fetal cells; and (f) analyzing at least one of the first nucleic acid sequence, the second nucleic acid sequence, the third nucleic acid sequence, the fourth nucleic acid sequence, or the fifth nucleic acid sequence to determine a likelihood that a fetus associated with an ex vivo population of fetal cells is susceptible to a genetic condition.

86. The method of claim 85, wherein at least one of the first nucleic acid sequence, the second nucleic acid sequence, the third nucleic acid sequence, the fourth nucleic acid sequence, or the fifth nucleic acid sequence comprises a DNA sequence.

87. The method of claim 85, wherein at least one of the first nucleic acid sequence, the second nucleic acid sequence, the third nucleic acid sequence, the fourth nucleic acid sequence, or the fifth nucleic acid sequence comprises a RNA sequence.

88. The method of any one of claims 85-87, wherein the ex vivo population of fetal cells is in a solution comprising a Artiodactyla serum.

89. The method of any one of claims 85-88, further comprising prior to (a)-(f), extracting an ex vivo population of maternal cells and the ex vivo population of fetal cells from a maternal blood sample to provide an extract, wherein the extract comprises the ex vivo population of maternal cells and the ex vivo population of fetal cells.

90. The method of claim 89, wherein the maternal blood sample is from a pregnant subject.

91. The method of claim 89, wherein the maternal blood sample is blood plasma.

92. The method of claim 89, wherein the maternal blood sample is blood serum.

93. The method of claim 89, wherein the maternal blood sample is buffy coat.

94. The method of claim 89, wherein the maternal blood sample comprises peripheral blood mononuclear white blood cells.

95. The method of claim 89, wherein the maternal blood sample comprises nucleated white blood cells.

96. The method of claim 89, wherein the maternal blood sample is whole blood.WSGR Docket No. 62193-701.601 97. The method of claim 89, further comprising enriching the ex vivo population of fetal cells in the extract from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

98. The method of claim 89, further comprising sorting the ex vivo population of fetal cells in the extract from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

99. The method of any one of claims 85-98, wherein prior to (a)-(f), the ex vivo population of fetal cells and an ex vivo population of maternal cells are extracted from a maternal blood sample to provide an extract, wherein the extract comprises the ex vivo population of fetal cells and an ex vivo population of maternal cells.

100. The method of claim 99, wherein the maternal blood sample is from a pregnant subject.

101. The method of claim 99, wherein the maternal blood sample is blood plasma.

102. The method of claim 99, wherein the maternal blood sample is blood serum.

103. The method of claim 99, wherein the maternal blood sample is buffy coat.

104. The method of claim 99, wherein the maternal blood sample comprises peripheral blood mononuclear white blood cells.

105. The method of claim 99, wherein the maternal blood sample comprises nucleated white blood cells.

106. The method of claim 99, wherein the maternal blood sample is whole blood.

107. The method of claim 99, wherein the ex vivo population of fetal cells in the extract is enriched from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

108. The method of claim 99, wherein the ex vivo population of fetal cells in the extract is sorted from the ex vivo population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

109. The method of any one of claims 85-108, further comprising prior to (a)-(f), incubating a blood sample comprising the ex vivo population of fetal cells and an ex vivo population ofWSGR Docket No. 62193-701.601 maternal cells with a first isolation agent and coupling the first isolation agent to a first biomarker in the ex vivo population of fetal cells.

110. The method of claim 109, further comprising sorting the ex vivo population of fetal cells based on a presence of the first isolation agent that couples to the first biomarker in the ex vivo population of fetal cells.

111. The method of claim 109, further comprising sorting the ex vivo population of fetal cells based on an absence of a second isolation agent that couples to a second biomarker in the ex vivo population maternal cells.

112. The method of claim 109, wherein the first isolation agent is a nucleic acid probe.

113. The method of claim 109, wherein the first isolation agent is a ligand.

114. The method of claim 109, wherein the first isolation agent couples to a cell surface biomarker.

115. The method of claim 109, wherein the first isolation agent couples to a soluble cell surface biomarker.

116. The method of claim 109, wherein the first isolation agent couples to an intracellular biomarker.

117. The method of claim 109, wherein the first isolation agent couples to a nucleic acid biomarker.

118. The method of any one of claims 85-117, wherein prior to (a)-(f), a blood sample comprising the ex vivo population of fetal cells and an ex vivo population of maternal cells is incubated with a first isolation agent that couples to a first biomarker in the ex vivo population of fetal cells.

119. The method of claim 118, wherein the ex vivo population of fetal cells is sorted based on a presence of the first isolation agent that couples to the first biomarker in the ex vivo population of fetal cells.

120. The method of claim 118, wherein the ex vivo population of fetal cells is sorted based on an absence of a second isolation agent that couples to a second biomarker in the ex vivo population maternal cells.WSGR Docket No. 62193-701.601 121. The method of claim 118, wherein the first isolation agent is a nucleic acid probe.

122. The method of claim 118, wherein the first isolation agent is a ligand.

123. The method of claim 118, wherein the first isolation agent couples to a cell surface biomarker.

124. The method of claim 118, wherein the first isolation agent couples to a soluble cell surface biomarker.

125. The method of clam 118, wherein the first isolation agent couples to an intracellular biomarker.

126. The method of claim 118, wherein the first isolation agent couples to a nucleic acid biomarker.

127. The method of any one of claims 85-126, further comprising prior to (a)-(f), incubating a blood sample comprising the ex vivo population of fetal cells and an ex vivo population of maternal cells with a second isolation agent, and coupling the second isolation agent to a second biomarker in the ex vivo population of maternal cells.

128. The method of claim 127, further comprising sorting the ex vivo population of maternal cells based on a presence of the second isolation agent that couples to the second biomarker in the ex vivo population of maternal cells.

129. The method of claim 127, further comprising sorting the ex vivo population of maternal cells based on an absence of a first isolation agent that couples to a first biomarker in the ex vivo population of fetal cells.

130. The method of claim 127, wherein the second isolation agent is a nucleic acid probe.

131. The method of claim 127, wherein the second isolation agent is a ligand.

132. The method of claim 127, wherein the second isolation agent couples to a cell surface biomarker.

133. The method of claim 127, wherein the second isolation agent couples to a soluble cell surface biomarker.WSGR Docket No. 62193-701.601 134. The method of claim 127, wherein the second isolation agent couples to an intracellular biomarker.

135. The method of claim 127, wherein the second isolation agent couples to a nucleic acid biomarker.

136. The method of any one of claims 85-135, wherein prior to (a)-(f), a blood sample comprising the ex vivo population of fetal cells and an ex vivo population of maternal cells is incubated with a second isolation agent that couples to a second biomarker in the ex vivo population of maternal cells.

137. The method of claim 136, wherein the ex vivo population of maternal cells is sorted based on a presence of the second isolation agent that couples to the second biomarker in the ex vivo population of maternal cells.

138. The method of claim 136, wherein the ex vivo population of maternal cells is sorted based on an absence of a first isolation agent that couples to a first biomarker in the ex vivo population of fetal cells.

139. The method of claim 136, wherein the second isolation agent is a nucleic acid probe.

140. The method of claim 136, wherein the second isolation agent is a ligand.

141. The method of claim 136, wherein the second isolation agent couples to a cell surface biomarker.

142. The method of claim 136, wherein the second isolation agent couples to a soluble cell surface biomarker.

143. The method of claim 136, wherein the second isolation agent couples to an intracellular biomarker.

144. The method of claim 136, wherein the second isolation agent couples to a nucleic acid biomarker.

145. The method of any one of claims 85-144, wherein the ex vivo population of fetal cells comprises early gestation trophoblasts.WSGR Docket No. 62193-701.601 146. The method of any one of claims 85-145, wherein the ex vivo population of fetal cells comprises later gestation trophoblasts.

147. The method of any one of claims 85-146, wherein the ex vivo population of fetal cells comprises erythroblasts.

148. The method of any one of claims 85-147, wherein the ex vivo population of fetal cells comprises megakaryocytes.

149. The method of any one of claims 85-148, wherein the ex vivo population of fetal cells comprises stromal cells.

150. The method of any one of claims 85-149, wherein the ex vivo population of fetal cells comprises endothelial cells.

151. The method of any one of claim 85-150, wherein the ex vivo population of fetal cells comprises lymphocytes.

152. The method of any one of claims 85-151, wherein the ex vivo population of fetal cells comprises nucleated red blood cells.

153. The method of any one of claims 85-152, wherein the ex vivo population of fetal cells comprises hematopoietic progenitor cells.

154. The method of any one of claims 85-153, wherein the ex vivo population of fetal cells comprises mesenchymal progenitor cells.

155. A method comprising: performing a single round of enrichment of a population of fetal cells from a population of maternal cells in a cell sample, wherein the single round of enrichment enriches the population of fetal cells from the cell sample to a level that provides a quantity of fetal nucleic acid that is sufficient for characterization as fetal nucleic acid rather than maternal nucleic acid.

156. The method of claim 155, wherein the single round of enrichment provides an enriched sample, wherein the level that provides the quantity of fetal nucleic acid that is sufficient for characterization as fetal nucleic acid rather than maternal nucleic acid is at least about 50% of fetal cells of the enriched sample.WSGR Docket No. 62193-701.601 157. The method of claim 156, comprising, performing one and no greater than one round of the enrichment.

158. The method of any one of claims 155-157, wherein the cell sample is a blood sample.

159. The method of claim 158, wherein the blood sample is blood plasma.

160. The method of claim 158, wherein the blood sample is blood serum.

161. The method of claim 158, wherein the blood sample is buffy coat.

162. The method of claim 158, wherein the maternal blood sample comprises peripheral blood mononuclear white blood cells.

163. The method of claim 158, wherein the maternal blood sample comprises nucleated white blood cells.

164. The method of claim 158, wherein the blood sample is whole blood.

165. The method of any one of claims 155-164, wherein the cell sample is from a human subject.

166. The method of any one of claims 155-165, further comprising prior to the performing, extracting the population of maternal cells and the population of fetal cells from the cell sample to provide an extract, wherein the extract comprises the population of maternal cells and the population of fetal cells.

167. The method of claim 166, further comprising sorting the population of fetal cells in the extract from the population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.

168. The method of any one of claims 155-167, wherein prior to the performing, the population of fetal cells and the population of maternal cells are extracted from the cell sample to provide an extract, wherein the extract comprises the population of maternal cells and the population of fetal cells.

169. The method of claim 168, wherein the population of fetal cells in the extract is sorted from the population of maternal cells in the extract to provide a sample enriched in fetal cells and depleted in maternal cells.WSGR Docket No. 62193-701.601 170. The method of any one of claims 155-169, wherein prior to the performing, the cell sample is incubated with a first isolation agent specific for a first biomarker in the population of fetal cells.

171. The method of claim 170, wherein the first isolation agent is a nucleic acid probe.

172. The method of claim 170, wherein the first isolation agent is a ligand.

173. The method of claim 170, wherein the first biomarker comprises a cell surface marker.

174. The method of claim 170, wherein the first biomarker comprises a soluble cell surface marker.

175. The method of claim 170, wherein the first biomarker comprises an intracellular biomarker.

176. The method of claim 170, wherein the first biomarker comprises a nucleic acid marker.

177. The method of any one of claims 155-176, further comprising prior to the performing, incubating the cell sample with a first isolation agent specific for a first biomarker in the population of fetal cells.

178. The method of claim 177, wherein the first isolation agent is a nucleic acid probe.

179. The method of claim 177, wherein the first isolation agent is a ligand.

180. The method of claim 177, wherein the first biomarker comprises a cell surface marker.

181. The method of claim 177, wherein the first biomarker comprises a soluble cell surface marker.

182. The method of claim 177, wherein the first biomarker comprises an intracellular biomarker.

183. The method of claim 177, wherein the first biomarker comprises a nucleic acid marker.

184. The method of any one of claims 155-183, wherein prior to the performing, the cell sample is incubated with a second isolation agent specific for a second biomarker in the population of maternal cells.

185. The method of claim 184, wherein the second isolation agent is a nucleic acid probe.

186. The method of claim 184, wherein the second isolation agent is a ligand.WSGR Docket No. 62193-701.601 187. The method of claim 184, wherein the second biomarker comprises a cell surface marker.

188. The method of claim 184, wherein the second biomarker comprises a soluble cell surface marker.

189. The method of claim 184, wherein the second biomarker comprises an intracellular biomarker.

190. The method of claim 184, wherein the second biomarker comprises a nucleic acid marker.

191. The method of any one of claims 155-190, further comprising prior to the performing, incubating the cell sample with a second isolation agent specific for a second biomarker in the population of maternal cells.

192. The method of claim 191, wherein the second isolation agent is a nucleic acid probe.

193. The method of claim 191, wherein the second isolation agent is a ligand.

194. The method of claim 191, wherein the second biomarker comprises a cell surface marker.

195. The method of claim 191, wherein the second biomarker comprises a soluble cell surface marker.

196. The method of claim 191, wherein the second biomarker comprises an intracellular biomarker.

197. The method of claim 191, wherein the second biomarker comprises a nucleic acid marker.

198. The method of any one of claims 155-197, further comprising, sequencing a nucleic acid sequence from the population of fetal cells.

199. A method comprising: a) extracting an ex vivo subpopulation of maternal cells and an ex vivo subpopulation of fetal cells from a maternal blood sample; b) after the extracting, enriching the ex vivo subpopulation of fetal cells from the ex vivo subpopulation of maternal cells; c) after the enriching, sorting the ex vivo subpopulation of fetal cells from the ex vivo subpopulation from maternal cells to provide a sample enriched in fetal cells and depleted in maternal cells; andWSGR Docket No. 62193-701.601 d) after the sorting, sequencing a nucleic acid from the ex vivo subpopulation of fetal cells.

200. A method comprising: a) contacting a maternal blood sample comprising an ex vivo subpopulation of fetal cells and an ex vivo subpopulation of maternal cells with an isolation agent that couples to a biomarker in the ex vivo subpopulation of fetal cells; b) after the contacting, incubating the isolation agent with the maternal blood sample; c) after the incubating, isolating the ex vivo subpopulation of fetal cells in the maternal blood sample from the ex vivo subpopulation of maternal cells in the maternal blood sample; and d) after the isolating, sequencing a nucleic acid from the ex vivo subpopulation of fetal cells.

201. A method comprising: a) contacting a maternal blood sample comprising an ex vivo subpopulation of fetal cells and an ex vivo subpopulation of maternal cells with an isolation agent that couples to a biomarker in the ex vivo subpopulation of fetal cells; b) after the contacting, incubating the isolation agent with the maternal blood sample; c) after the incubating, isolating the ex vivo subpopulation of fetal cells in the maternal blood sample from the ex vivo subpopulation of maternal cells in the maternal blood sample; d) after the isolating, sorting the ex vivo subpopulation of fetal cells from the ex vivo subpopulation of maternal cells; and e) after the sorting, sequencing a nucleic acid from the ex vivo subpopulation of fetal cells.

202. A method comprising: incubating a population of cells from a sample of a subject with a first isolation agent, a second isolation agent, a third isolation agent, a fourth isolation agent, a fifth isolation agent, and a sixth isolation agent, wherein the population of cells comprises a subpopulation of fetal cells and a subpopulation of maternal cells, wherein the first isolation agent is specific for a first biomarker, the second isolation agent is specific for a second biomarker, the third isolation agent is specific for a third biomarker, the fourth isolation agent is specific for a fourth biomarker, and the fifth isolation agent is specific for a fifth biomarker inWSGR Docket No. 62193-701.601 the subpopulation of fetal cells, and wherein the sixth isolation agent is specific for a sixth biomarker in the subpopulation of maternal cells.

203. The method of claim 202, wherein the subject is a human subject.

204. The method of claim 202 or claim 203, wherein the sample is a blood sample.

205. The method of claim 204, wherein the blood sample is blood plasma.

206. The method of claim 204, wherein the blood sample is blood serum.

207. The method of claim 204, wherein the blood sample is buffy coat.

208. The method of claim 204, wherein the maternal blood sample comprises peripheral blood mononuclear white blood cells.

209. The method of claim 204, wherein the maternal blood sample comprises nucleated white blood cells.

210. The method of claim 204, wherein the blood sample is whole blood.

211. The method of any one of claims 202-210, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is a nucleic acid probe.

212. The method of any one of claims 202-211, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is a ligand.

213. The method of any one of claims 202-212, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a cell surface marker.

214. The method of any one of claims 202-213, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a soluble cell surface marker.WSGR Docket No. 62193-701.601 215. The method of any one of claims 202-214, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for an intracellular biomarker.

216. The method of any one of claims 202-215, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a nucleic acid marker.

217. The method of any one of claims 202-216, further comprising after the incubating, enriching the subpopulation of fetal cells from the subpopulation of maternal cells.

218. The method of any one of claims 202-217, wherein after the incubating, the subpopulation of fetal cells is enriched from the subpopulation of maternal cells.

219. A method comprising: incubating a population of cells with a first isolation agent, a second isolation agent, a third isolation agent, a fourth isolation agent, a fifth isolation agent, and a sixth isolation agent, wherein the population of cells comprises a subpopulation of fetal cells and a subpopulation of maternal cells from a sample of a subject, and wherein the first isolation agent is specific for a first biomarker in the subpopulation of fetal cells, the second isolation agent is specific for a second biomarker in the subpopulation of fetal cells, the third isolation agent is specific for a third biomarker in the subpopulation of fetal cells, the fourth isolation agent is specific for a fourth biomarker in the subpopulation of maternal cells, the fifth isolation agent is specific for a fifth biomarker in the subpopulation of maternal cells, and the sixth isolation agent is specific for a sixth biomarker in the subpopulation of maternal cells.

220. The method of claim 219, wherein the subject is a human subject.

221. The method of claim 219 or claim 220, wherein the sample is a blood sample.

222. The method of claim 221, wherein the blood sample is blood plasma.

223. The method of claim 221, wherein the blood sample is blood serum.

224. The method of claim 221, wherein the blood sample is buffy coat.

225. The method of claim 221, wherein the maternal blood sample comprises peripheral blood mononuclear white blood cells.WSGR Docket No. 62193-701.601 226. The method of claim 221, wherein the maternal blood sample comprises nucleated white blood cells.

227. The method of claim 221, wherein the blood sample is whole blood.

228. The method of any one of claims 219-227, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is a nucleic acid probe.

229. The method of any one of claims 219-228, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is a ligand.

230. The method of any one of claims 219-229, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a cell surface marker.

231. The method of any one of claims 219-230, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a soluble cell surface marker.

232. The method of any one of claims 219-231, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for an intracellular biomarker.

233. The method of any one of claims 219-232, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, and the sixth isolation agent is specific for a nucleic acid marker.

234. The method of any one of claims 219-233, further comprising after the incubating, enriching the subpopulation of fetal cells from the subpopulation of maternal cells.

235. The method of any one of claims 219-234, wherein after the incubating, the subpopulation of fetal cells is enriched from the subpopulation of maternal cells.

236. A method comprising: incubating a population of cells with a first isolation agent, a second isolation agent, a third isolation agent, a fourth isolation agent, a fifth isolation agent, a sixthWSGR Docket No. 62193-701.601 isolation agent, and a seventh isolation agent, wherein the population of cells comprises a subpopulation of fetal cells and a subpopulation of maternal cells from a sample of a subject, wherein the first isolation agent is specific for a first biomarker, the second isolation agent is specific for a second biomarker, the third isolation agent is specific for a third biomarker, the fourth isolation agent is specific for a fourth biomarker, the fifth isolation agent is specific for a fifth biomarker, and the sixth isolation agent is specific for a sixth biomarker in the subpopulation of maternal cells, and wherein the seventh isolation agent is specific for a seventh biomarker in the subpopulation of fetal cells.

237. The method of claim 236, wherein the subject is a human subject.

238. The method of claim 236 or claim 237, wherein the sample is a blood sample.

239. The method of claim 238, wherein the blood sample is blood plasma.

240. The method of claim 238, wherein the blood sample is blood serum.

241. The method of claim 238, wherein the blood sample is buffy coat.

242. The method of claim 238, wherein the maternal blood sample comprises peripheral blood mononuclear white blood cells.

243. The method of claim 238, wherein the maternal blood sample comprises nucleated white blood cells.

244. The method of claim 238, wherein the blood sample is whole blood.

245. The method of any one of claims 236-244, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent comprises a nucleic acid probe.

246. The method of any one of claims 236-245, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the f ourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent comprises a ligand.

247. The method of any one of claims 236-246, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent is specific for a cell surface marker.WSGR Docket No. 62193-701.601 248. The method of any one of claims 236-247, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent is specific for a soluble cell surface marker.

249. The method of any one of claims 236-248, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent is specific for an intracellula r biomarker.

250. The method of any one of claims 236-249, wherein one of the first isolation agent, the second isolation agent, the third isolation agent, the fourth isolation agent, the fifth isolation agent, the sixth isolation agent, and the seventh isolation agent is specific for a nucleic acid marker.

251. The method of any one of claims 236-250, further comprising after the incubating, enriching the subpopulation of fetal cells from the subpopulation of maternal cells.

252. The method of any one of claims 236-251, wherein after the incubating, the subpopulation of fetal cells is enriched from the subpopulation of maternal cells.

Images