66 results about "Cell lineage" patented technology

The invention provides a cell development process dynamic modeling method and device based on time sequence single cell transcriptome data and a medium, and relates to the crossing field of bioinformatics and computational biology. The method comprises the following steps: constructing a Shenchang differential equation learning framework; adjusting parameters of the single cell development state change model based on the Shenxuan differential equation learning framework so as to construct a population cell development state change model; obtaining a cell specific gene regulation network and a population cell gene regulation network based on the population cell development state change model so as to predict occurrence opportunity of cell lineage differentiation and a molecular decision mechanism of cell differentiation; therefore, the problems of incomplete modeling mechanism, insufficient noise processing and lack of energy principle in the existing cell development process are solved.

The invention discloses a cell fate distinguishing and differentiation prediction method based on a three-dimensional form, and belongs to the technical field of cell fate distinguishing and differentiation prediction. In order to solve the problem that cell fate differentiation is difficult to distinguish and predict under a small sample size, the method comprises the following steps: acquiring a living biological sample, and imaging by using a three-dimensional delay microscope to obtain cell three-dimensional image data; segmenting the cell membrane boundary and extracting geometric and high-order three-dimensional morphological characteristic parameters; labeling cell fate and differentiation states based on means such as cell lineage tracking or gene fluorescence labeling; and finally, determining an optimal morphological index and a minimum sample size through single-side double-sample t test, and applying the optimal morphological index and the minimum sample size to two new cell populations and outputting a cell fate distinguishing and differentiation prediction result. The method can realize cross-species, low-loss, low-cost and high-precision cell fate differentiation and differentiation prediction, and is suitable for development research, stem cell culture and disease diagnosis.

The invention discloses a cell lineage tracking system based on synNotch and CRISPR / Cas9 bar code technology and application thereof, the cell lineage tracking system comprises a system mGFP ligand vector for constructing Sender ligand cells and a system for constructing Receiver recipient cells, and the system comprises a synNotch receptor-rtTA fusion vector, a TetO-Cas9 expression vector, a gRNA expression vector and a Target vector library containing a Barcode sequence. A synNotch system and a CRISPR / Cas9 gene editing technology are combined with a bar code strategy, a unique bar code is generated, the contact sequence between cells is recorded, the method is compatible with a single-cell sequencing technology, transcription information of the cells is provided, long-term tracking of the contact history between the cells is achieved, the method can be used for tracking interaction and functions between the cells in a tumor microenvironment, and the application prospect is wide. Through long-term tracking and functional analysis of interaction between macrophages, especially macrophages and tumor cells, the accuracy of interaction analysis is remarkably improved, and a more accurate target spot is provided for targeted therapy.

The present invention relates to the new use of an exosome secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and in particular relates to the use of the exosome in the preparation of a drug for treating liver diseases. The exosome is secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and the hepatocyte lineage cells are hepatic progenitor cells and / or mature hepatocyte-like cells.

A method for in situ differentiation of stem cells within a cell culture bioreactor is provided. The method includes providing a bioreactor vessel having a cell culture chamber, the cell culture chamber having an inlet for introducing fluid into the cell culture chamber and an outlet for introducing fluid out of the cell culture chamber. The bioreactor vessel also includes a cell substrate disposed within the cell culture chamber for culturing cells on the cell substrate. The method includes seeding undifferentiated stem cells onto the cell substrate within the cell culture chamber and perfusing the cell culture chamber with a differentiation medium to promote differentiation of the undifferentiated stem cells into specific cell lineages, thereby causing the undifferentiated stem cells to become differentiated cells.

A method of generating cells of the T cell lineage is provided that involves culturing a sample comprising stem cells or progenitor cells with an engineered Notch ligand conjugated to a suspension support and isolating cells of the T cell lineage. In one embodiment, the cells of the T-cell lineage are progenitor T cells or mature T cells. Compositions, kits and uses thereof are also provided.

Provides herein is a method for identifying the specific cell lineage of cells in culture comprising the steps of determining from the nucleic acid molecules isolated from said recombinant cells in culture the presence of polymorphisms or SNPs at at least 5 different positions within at least five genes contained in said nucleic acid molecules, obtaining a genetic profile from the determination of the previous step, and identifying the cell lineage of said cells in culture from said genetic profile, and wherein the recombinant cells produce a recombinant protein.

Disclosed are methods and compositions for generating B lineage cells, such as plasmablasts, plasma cells, and protein-producing or protein-secreting B cells. The methods may relate to stage-specific differentiation from stem cells or stem-cell derived hematopoietic progenitors through one or more intermediates to B lineage cells, such as plasmablasts, plasma cells, and protein-producing or protein-secreting B cells.

The present application relates to a kind of pluripotent stem cell co-differentiation method for generating vascularized heart organoids, the method comprises: (1) in iPSCs, obtain iEC-iPSCs by transferring exogenous gene Tet 3G and ETV2;(2) the iEC-iPSCs in step (1) are mixed with iPSCs, add culture medium I, prepare blastula after culture;(3) the blastula in step (2) is induced by using the heart organoid differentiation method combined with doxycycline, and vascularized heart organoids are prepared after differentiation culture.The present application uses Tet-on 3G inducible expression system, promotes the differentiation of part of cells in heart organoids to endothelial cell lineage by regulating the expression of ETV2 gene in part of iPSCs in blastula, and then forms vascularized heart organoids.

The application discloses a human vagina micro-ecological in-vitro model and a construction method and application thereof, and relates to the technical field of medicines.The application discloses a human vagina micro-ecological in-vitro model and a construction method and application thereof.Firstly, a mixed cell group of the vaginal epithelium containing a complete cell lineage and capable of long-term subculture is successfully established, a technical bottleneck that primary vaginal epithelial cells are difficult to expand is solved, and ideal seed cells are provided for model construction.Secondly, a highly-bionic vaginal mucosa model having a tight connection structure, rich layers and an orderly arrangement is constructed, and the physiological barrier and immune regulation functions of the vaginal mucosa are truly simulated.Finally, by introducing lactobacillus symbiosis and confirming the response to estrogen, a multi-element in-vitro research model integrating the anatomical structure of the vagina, resident flora, local immunity and endocrine regulation is constructed, and a reliable technical platform is provided for in-depth exploration of the mechanism of vaginal micro-ecological disorder and efficient screening of related drugs.

The invention discloses application of a series of extremely early stem cells or differentiated cell lineages thereof in preparation and detection of influenza viruses and drug screening, and belongs to the field of cell application. Specifically, trophoblast stem cells (TSC) and amnion-like stem cells (AME) obtained by extremely early cell induced differentiation and screening are used for influenza virus infection or screening test. Experiments prove that extraembryonic differentiation cell models such as trophoblast stem cells, amniotic stem cells and / or differentiated cell lineages thereof are highly susceptible to influenza viruses, and have important application values in the aspects of efficient preparation and detection of viruses and variants thereof, preparation of preventive vaccines and screening of therapeutic drugs. The method not only aims at human-derived influenza viruses and variants thereof, but also has important significance on prevention and treatment of animal influenza and variants thereof.

The present disclosure provides bovine animals and methods to create recipient bovine animals for spermatogenic stem cell transplantation / complementation through modulation of the NANOS2 gene. In one embodiment genome editing was used to create bovine animals with insertions or deletions (indels) that inactivate or otherwise modulate NANOS2 gene activity so that resulting male bovines lack functional germ cells yet retain functional testicular somatic cells, and bovine females are fertile. These bovine males can then be transplanted or complemented with donor cells capable of giving rise to cells of the spermatogenic lineage, or spermatogenic stem cells and used for breeding.

The invention relates to the field of bioinformatics, and provides a single cell pedigree tracing method. According to the technical scheme, single cell transcriptome sequencing is combined with Bulk DNA targeted sequencing, so that the cost of pure single cell transcriptome sequencing is reduced, and the accuracy of single cell labeling is ensured through a subsequent filtering method of reserving a unique bar code.

Provided herein are methods, compositions, systems, kits and uses for integrated, simultaneous spatial multiomic analyses of tissues and samples including chromatin accessibility epigenomic analysis, RNA expression transcriptomic analysis, cell cluster and cell lineage analysis, transcription factor motif analysis, extrachromosomal DNA analysis and mitochondrial DNA analysis.

The application discloses a high-precision and high-coverage pedigree tree tracking method, and relates to the technical field of cell pedigree tracking, and specifically comprises the following steps: selecting single-direction RNA1, single-direction RNA2, single-direction RNA3 and single-direction RNA4 which are completely different in sequence as backups, and selecting single-cell clone amplification to prepare a pedigree barcode with 13 editing sites as a backup; and adopting a cell pedigree barcode technology to match the single-direction RNA1, the single-direction RNA2, the single-direction RNA3 and the single-direction RNA4 with the 13 editing sites of the pedigree barcode respectively. The high-precision and high-coverage pedigree tree tracking method increases the average expression amount of the pedigree barcode in all cells, reduces the speed of consumption of the editing sites, increases the traceable time, simultaneously reduces the proportion of cross-site deletion mutations, and obtains a high proportion of single leaves of a cell pedigree tree, so that a pedigree tree determination method with a single-cell level resolution is realized, and the mortality of cells in clone amplification and the loss rate in cell digestion are reduced.

Animal cells, notably adult fibroblasts, are advantageously reprogrammed in direct lineage reprogramming methods using defined factors to produce proliferative and multipotent induced cardiac progenitor cells (iCPC). The iCPC thus produced can be differentiated under suitable differentiation conditions to cardiac lineage cells including cardiomyocytes, smooth muscle cells, and endothelial cells, as evidenced by expression of lineage specific markers. Sets of factors effective in combination to reprogram the fibroblasts can include a set that includes some or all of 5 factors (Mespl, Baf60c, Nkx2.5, Gata4, Tbx5), a set that includes some or all of 11 factors (Mespl, Mesp2, Gata4, Gata6, Baf60c, SRF, Isll, Nkx2.5, Irx4, Tbx5, Tbx20), a set that includes some or all of 18 factors (T, Mespl, Mesp2, Tbx5, Tbx20, Isll, Gata4, Gata6, Irx4, Nkx2.5, Handl, Hand2, Tbx20, Tbx18, Tip60, Baf60c, SRF, Hey2), and a set that includes some or all of 22 factors (T, Mespl, Mesp2, Tbx5, Tbx20, Isll, Gata4, Gata6, Irx4, Nkx2.5, Hand1, Hand2, Tbx20, Tbx18, Tip60, Baf60c, SRF, Hey2, Oct4, Klf4, Sox2, L-myc).

There is provided a method of generating T cell lineage cells, the method comprising (a) culturing a sample comprising stem cells or progenitor cells with a soluble multispecific ligand comprising a Notch ligand and a functional protein partner (FPP) that binds to a surface molecule on the stem cells or progenitor cells, and (b) isolating the T cell lineage cells. In one embodiment, the T cell lineage cells are progenitor T cells or mature T cells. Multispecific ligands, compositions, kits and uses thereof are also provided.

This invention discloses a composition for the differentiation of mesenchymal stem cells (ADSCs) into keratinocytes and its application. The composition comprises: a Wnt signaling pathway activator, a TGF-β signaling pathway inhibitor, a bone morphogenetic protein pathway activator, and at least one of retinoic acid and a retinoic acid agonist. The small molecule composition provided by this invention is meticulously designed to gradually guide the fate transition of ADSCs by mimicking key signaling events during embryonic development. This strategy is not a simple compound screening, but a rational design based on a deep understanding of developmental biology, achieving precise regulation of cell lineages in vitro by reproducing the natural developmental process. The composition of this invention is not a simple superposition of components, but rather achieves cell fate transition by precisely regulating the balance of signaling pathway networks.

The present specification provides: a method of isolation of a pure culture of vascular endothelial cells, the method capable of isolating homogeneous endothelial cells adhered to a matrix for a specific time in a cell line of an endothelial cell lineage differentiated from human pluripotent stem cells; a medium for maintaining characteristics of vascular endothelial cells, comprising high-purity vascular endothelial cells isolated through the method, 4 ng / ml to 6 ng / ml of FGF2, 5 ng / ml to 10 ng / ml of EGF, 10 ng / ml to 30 ng / ml of VEGF-A, 20 ng / ml to 50 ng / ml of ascorbic acid, and DMEM / F-12 as active ingredients; and a culture method comprising same.

The invention relates to a cell fate determinant identification method and a related device, and belongs to the technical field of bioinformatics, the cell fate determinant identification method comprises the following steps: determining expression quantities of transcription factors and target genes in cells in single cell transcriptome sequencing data, and constructing a cell lineage; constructing a gene regulation and control model based on the relationship between the expression quantity of each transcription factor in each cell cluster in the cell lineage in the cells and the expression quantity of each target gene in the cells; constructing a target function according to the gene regulation model precision of each cell cluster, the gene regulation model sparseness of each cell cluster and the gene regulation model similarity between the adjacent cell clusters, and determining a regulation coefficient of a transcription factor in each cell cluster to a target gene based on the target function; and determining a differential transcription factor between adjacent cell clusters based on the regulation coefficient, and determining a cell fate determinant based on the differential transcription factor. According to the invention, the cell fate determinant can be accurately identified.

The invention relates to a detection kit for a peripheral blood complete immune cell lineage and a door analysis model thereof. The detection kit comprises a monoclonal antibody which is combined with a peripheral mononuclear cell antigen and is marked by 33 rare metal elements. The detection method comprises the following steps: (1) a PBMC (peripheral blood mononuclear cell) acquisition unit: carrying out PBMC separation on a human peripheral blood sample; (2) a living cell bar code marking unit: performing dyeing marking on a plurality of PBMC samples by using a CD45 antibody combination marked by three different metal elements; (3) a sample dyeing unit: mixing a plurality of samples marked by bar codes, and performing dyeing incubation by using prepared 28 metal marked antibodies mix; and (4) a data acquisition unit: performing detection by using a mass spectrometer to obtain original data of the mixed samples. According to the method, rapid and accurate analysis of multi-sample high-dimensional data can be realized, the consumption of precious samples is remarkably reduced, and the cost benefit is considered.

Provided herein are compositions and methods for method for inducing intermediate macrophages in a subject, the method comprising: administering to the subject a therapeutically effective amount of one or more compositions that comprise a compound of formula (I) or stereoisomer, enantiomer, tautomer or a pharmaceutically acceptable salt thereof (I) wherein the compound promotes monocyte differentiation into an antigen-presenting cell (APC)- specific intermediate macrophage lineage.

A normal set of cells is characterized using flow cytometry. A centroid and radius are defined for a set of clusters in an n-dimensional space corresponding to a normal maturation for a cell lineage in the normal set of cells. A test set of cells is characterized using flow cytometry and the characterization is compared to the defined set of clusters. Support Vector Machine (SVM) subroutines are employed to identify reference populations of interest by generating multidimensional boundary definitions. These boundary definitions may be used to identify reference populations to use in defining or refining a centroid line or a radius or radii defining a set of normal clusters, and to characterize and compare a test set of cells to the defined set of normal clusters.

The invention discloses a DNA structure for saccharomyces cerevisiae single-cell barcode labeling and chromatin state conjoint analysis and application thereof, and belongs to the technical field of molecular biology and single-cell multi-omics analysis. From the 5'end to the 3 'end, a fluorescent protein gene, a Tn5ME structure, a reverse transcription primer binding region, a CYC1 terminator fragment and a linearized shuttle plasmid skeleton are subjected to homologous recombination, and a pCM carrier is obtained; after NdeI / BamHI enzyme digestion, oligonucleotides with random sequences, which are subjected to annealing, amplification, Klenow filling-in and enzyme digestion, are connected, and transformation extraction is carried out, so that the DNA structure for saccharomyces cerevisiae single-cell barcode labeling and chromatin state combined analysis is obtained. On the basis of the constructed DNA structure, ATAC-seq and a bar code reverse transcription method are combined, important data related to the transcription state and chromatin state of a single cell can be obtained at the same time through bar code information, single cell pedigree tracking of the saccharomyces cerevisiae is achieved, and research on single cell epigenomics of the saccharomyces cerevisiae and other microorganisms is promoted.

The present disclosure provides a method for generating a population of T cell lineages from T cell progenitor cells. The method comprises culturing the progenitor cell in the presence of a Notch signaling ligand, such as delta-like 4 (DL4), provided on a surface area of at least 7 square centimeter / ml culture volume (7cm2 / mL). Provided are populations of cells produced using the methods and methods of using the populations of cells. Further provided is a method of differentiating a population of T cell progenitor cells enriched for CD4-CD8 + TCR [gamma] [delta] + cells, the method comprising culturing the T cell progenitor cells in the presence of a Notch signaling ligand, such as DL4, provided on a surface area of 0.78 to 4.7 cm2 / mL.

The invention provides a construction method of an embryo-like model, which comprises the following steps: culturing primitive state stem cells in vitro or injecting the primitive state stem cells into an immunodeficient animal body for differentiation to obtain the embryo-like model. According to the method, in the process of preparing the embryo-like model, cell factors and small molecular pathway interference factors do not need to be added, gene editing is not needed, only a single starting cell type is adopted, and the method has the advantages of being simple and high in repeatability, facilitates large-scale preparation and application, and has wide application prospects. And the prepared embryo-like model has all embryo external cell lineages and all embryo internal cell lineages except for trophoderm, and can simulate the development process from an implanted embryo of a Carbonate-resistant group 3 stage to a Carbonate-resistant group 9 stage nerve embryo, so that the embryo-like model can be used for preparing various progenitor cell lineages, modeling diseases, screening drugs and carrying out scientific research.

The invention belongs to the technical field of molecular biology, and particularly relates to a self-guiding type stpegRNA, a recombinant plasmid containing the self-guiding type stpegRNA and a CRISPR editing method for cell lineage tracking. The core of the invention is a DNA sequence, after the DNA sequence is transcribed into pegRNA, editing and self-extension of the DNA can be re-identified and mediated, and the sequence is characterized in that 5 '-3' sequences at the head end and the tail end of the pegRNA are basically consistent, and a pegRNA skeleton is subjected to homing modification. The development of the Infinite Printer marks the new era of the CRISPR pedigree tracking technology from limited recording to infinite analysis. Through fundamental reconstruction of an editing mode, collaborative breakthrough of capacity, resolution and safety is realized, and a brand new normal form is provided for research of developmental biology, oncology and regenerative medicine. In the future, along with the improvement of editing efficiency and the innovation of a calculation algorithm, the system is expected to become a standard tool for analyzing a complex biological system, and plays a reform role in the fields of personalized medical treatment, early diagnosis of diseases and the like.

The invention discloses a device, a reagent and a kit for detecting B19V infection and a method for identifying B19V multilineage cell infection, and belongs to the field of virus detection. The technical problem to be solved by the invention is to evaluate B19V infection conditions of different cell lineages in the same sample. The reagent for detecting B19V infection disclosed by the invention contains a B19V probe and an antibody for marking a cell lineage, the sequence of the B19V probe is as shown in SEQ ID No.1, and the antibody for marking the cell lineage is a CD34 antibody, a CD38 antibody, a CD4 antibody, a CD8 antibody and a CD56 antibody. The reagent and the kit can be used for simultaneously detecting the B19V infection condition of multi-lineage cells in the same sample, distinguishing the B19V infection conditions of different cell lineages in the same sample, and realizing accurate positioning of B19V positive cells.

A system and method for analyzing a sputum sample from a subject suspected of having lung cancer comprising obtaining a plurality of cells from the sputum sample from the subject, marking the plurality of cells with i) a plurality of cell lineage specific marker compositions, ii) a cell viability composition and iii) a tetra (4-carboxyphenyl) porphyrin (TCPP) composition; analyzing with the flow cytometer the plurality of cells marked with i-iii to obtain a subpopulation selected for cell size from the plurality of cells based upon an automatically selected bead size exclusion gate; from the cell size selected subpopulation, selecting a viable singlet population of cells using an automated non-debris gate and an automated singlets gate; from the viable singlet population of cells, obtaining flow cytometer values based upon the plurality of cell lineage specific marker compositions, the viability marker and the TCPP marker; applying a trained classifier to meta data from the subject and the flow cytometric values obtained; and generating, based upon the application of the trained classifier, a classification for the sputum sample wherein the classification is selected from a plurality of classification options comprising cancer and non-cancer.