171 results about "Trophoblast cell" patented technology

The present invention relates to a non-invasive method of retrieving and identifying cells particularly fetal cells and trophoblastic cells. The invention includes methods for use of the cells for identifying chromosomal abnormalities and mutations particularly for prenatal diagnosis by performing genetic diagnosis for chromosomal and single gene disorders. The invention also includes methods of confirming cells of fetal origin.

The invention belongs to the technical field of medicine, and specifically relates to a primary tumor cell culture medium, culture method and application. The primary tumor cell culture method provided by the invention comprises the steps as follows: preparing a primary cell culture medium which comprises the following components: hydrocortisone, EGF, Insulin, a ROCK inhibitor, but not contains cholera toxin, and is an improvement of the existing primary cell culture medium; culturing tumor tissue epithelial cells on laid-out trophoblast cells by using the primary cell culture medium, and enabling the tumor tissue epithelial cells to proliferate rapidly under the combined action of growth factors secreted by the trophoblast cells and nutrient factors contained in the culture medium; and digesting and sub-culturing the tumor tissue epithelial cells when the tumor tissue epithelial cells grow to a cell density of about 80% to 90%. A convenient primary cell culture method is used to obtain immortalized cells possessing the biological characteristics of a patient's own tumor, and the problem of immortalization of the primary culture of tumor cells is solved, thereby realizing personalized treatment for the patient.

The invention discloses a method for in-vitro amplification of NK cells and the NK cells obtained by the same. The method comprises the following steps: taking peripheral blood mononuclear cells (PBMC) separated out from peripheral blood as seed cells, adding the cells into a culture bottle pre-coated with CD16Mab when induced culture begins and enlarged culture is carried out every time, performing continuous stimulation on the cells, and finally obtaining a large amount of high-purity NK cells. Under the situation that the stimulation is performed without using trophoblast cells, the large amount of high-purity NK cells is finally obtained through the method of performing continuous stimulation by coating the culture bottle with CD16Mab, and the use amount of cell factors is reduced.

The invention discloses an embryo pregnancy result prediction device based on multimodality, and belongs to the field of medical artificial intelligence. Firstly, images of embryos developed to blastocyst stage after in vitro fertilization and corresponding pregnancy results are obtained, three pictures of blastocyst, inner cell mass and trophoblast cells of embryos are obtained, and the pregnancyresults are labeled as labels, and data are labeled as raw data. Then the image is smoothed with Gaussian kernel function to remove part of the noise, and then the image is normalized. The image is then data-augmented as input data. Multimodal method is used to fuse the images of the three images so that the input image contains three evaluation features. the fused image is passed to ResNet-50 for training, that network is optimized according to the target tag, and and is iterated until the training is complete. With the model, three images can be taken before embryo transfer to predict the pregnancy outcome, and the embryo with high success rate can be selected according to the output results, which can improve the final pregnancy success rate.

The invention discloses a novel ips cell inducing method and provides application of inducing genes Oct3 / 4, Klf4, Sox2, P53, L-Myc, Lin28, RARg and Lrh-1 in inducing isolated mammal somatic cells to generate iPS multipotent stem cells. By an iPS cell establishing method, a method for inducing human iPS stem cells free of exogenous serum, trophoblast cells and animal source is established on the basis of no insertion. The method shortens reprogramming time of the iPS multipotent stem cells and improves cloning generation efficiency, and obtained iPS cells are higher in dryness.

The invention discloses a serum-free culture medium used for culturing and amplifying skin keratinocytes in vitro. It adds amino acid, vitamin, hormone, growth gene, trace elements, and antioxidant, thus able to accelerate cell breeding and delay consenescence of the keratinocytes and makes them amplified in vitro by more times. Using the culture medium to make primary culture and subculture needs no trophoblast cells and no extracellular substrates like collagen, etc prepaved at the bottom of the culture bottle to help the keratinocytes adhere to the wall and grow.

The first method to cause a culture of human and other primate stem cells to directly and uniformly differentiate into a committed cell lineage is disclosed. Treatment of primate stem cells with a single protein trophoblast induction factor causes the cells to transform into human trophoblast cells, the precursor cells of the placenta. Several protein factors including bone morphogenic protein 4 (BMP4), BMP2, BMP7, and growth and differentiation factor 5 can serve as trophoblast-inducting factors.

The invention provides a method for culturing human induced pluripotent stem cells (iPSCs) by using human bone marrow mesenchymal stem cells as a trophoblast. The method comprises the following steps of: obtaining the human iPSCs from child foreskin fibroblasts by using third to fifth generation human bone marrow mesenchymal stem cells obtained through subculture and culture after thawing from low-temperature freezing, performing amplification culture, and inoculating into trophoblast cells to obtain the human iPSCs. In the method, the human iPSCs are cultured by using the human bone marrow mesenchymal stem cells as the trophoblast cells instead of mouse embryonic fibroblasts in the conventional method, so that the pollution of heterologous cells in a culture system is reduced, that the culture system can make the human iPSCs amplified in vitro is proved, biological characteristics and multipotency of the human iPSCs are maintained for a long time, and the possibility of clinical application of the human iPSCs is provided.

The invention relates to an in-vitro pure culture method for natural killer cells. The method includes the following steps that peripheral blood is collected, single karyocyte is separated, magnetic beads combined with specific antibodies are adopted to remove the NK cells, and NK cell populations are obtained; the NK cell populations are put into a culture bottle coated with multiple antibodies,and are subjected to activation culture with a complete medium containing IL-2, IL-7, IL-15, anti-CD16, OK432 and inactivated autologous serum, and inactivated NK cell populations are obtained; the inactivated NK cell populations are put into a culture bottle free of antibody coating and continue to carry out multiplication culture, and the natural killer cells are obtained. Any heterologous serumingredients and trophoblast cells are not used in the in-vitro pure culture method, and high amplification efficiency can be wholly obtained; meanwhile, the purity of the amplified NK cells reaches 90% or above, in-vitro efficient pure culture is achieved, and the clinical application requirement is conveniently met.

The invention relates to a method for in vitro multiplication culture of NK cells. The method comprises coating a culture bottle with heparin sodium, CD16 and human immunoglobulin, collecting peripheral blood, inactivating plasma, separating mononuclear cells, using a serum-free medium (containing autologous plasma, IL-2, IL-15 and IL-18) to culture multiplication cells, harvesting and detecting the cells and the like. Trophoblast cells or magnetic beads are not used for sorting and purifying, and the method has simple operation, high safety, good amplification effect and high clinical application value.

The present invention provides antibodies that specifically bind to trophoblast cell-surface antigen-2 (Trop-2). The invention further provides antibody conjugates comprising such antibodies, antibody encoding nucleic acids, and methods of obtaining such antibodies. The invention further relates to therapeutic methods for use of these antibodies and Trop-2 antibody conjugates for the treatment of a condition associated with Trop-2 expression (e.g., cancer), such as colon, esophageal, gastric, head and neck, lung, ovarian, or pancreatic cancer.

The invention relates to a stem cell culture medium which contains certain chemical components as follows: water-soluble non-polyelectrolyte polymers, inorganic salts, vitamins, amino acids, glucose, transferrin, insulin or a substitute with the same functions as the insulin (the IGF-1 / 2), and a fibroblast growth factor. The stem cell culture medium containing the certain chemical components is free of additives from an animal source, namely various animal products, trophoblast cells and conditioned mediums, and has the advantages that cell differentiation resulted from non-absolute purification of the traditional culture medium blood products can be avoided; a totipotential stem cell culture medium can be provided to effectively increase human totipotential stem cells (including the hESC and the hiPSC) in vitro, achieve a remarkable effect, and substitute for the traditional culture mediums from an animal source; due to the adoption of the stem cell culture medium stem cell culture medium, the quality stability of each product batch can be ensured effectively to facilitate the product standardization.

The invention discloses an in-vitro expansion method, a kit and an application of umbilical cord blood NK (nature killer) cells. The in-vitro expansion method of the umbilical cord blood NK cells comprises the processes of activated culture and enrichment culture of the umbilical cord blood NK cells from isolated cord blood mononuclear cells. Compared with existing methods for isolating and expanding the umbilical cord blood NK cells, the in-vitro expansion method has the advantages that cell sorting is needless, trophoblast cells are needless, operation is simple, universality is good, cost is low, adopted reagents do not contain components of animal origin, safety is good, the yield of the NK cells is high, purity of the obtained cord blood NK cells is up to 90% or higher, the total number of the cells can be up to 10<10-11> / part of cord blood, and the killing capacity for tumor cells is high, thereby playing a role in preparation of tumor immunotherapy drugs and tumor immunotherapy.

The invention provides a method for producing mesenchymal stem cells through induction of human pluripotent stem cells (hPSC). The method comprises the following steps: performing continuous treatmenton the hPSC by a low sugar medium on the basis of a PSC culture system without mouse trophoblast, coating the culture system by gelatin, performing pressure screening on the cells, and performing continuous unicellular passage on the cells through trypsin, and successfully optimizing a simple and high-efficiency method for differentiating the hPSC ino hMSC. The method disclosed by the invention can be used for simultaneously producing lots of functional hPSC-derived hMSC. Meanwhile, the differentiation process is less in time, participation of trophoblast cells, embryoid, flow cytometry, small molecule compounds and special culture conditions is avoided, the operation is simple, and the cost is low.

The invention discloses a conditional Cas9 expression induced swine trophoblastic cell line and an establishment method and application thereof. The method includes: subjecting a lentiviral vector, for controlling Cas9 gene expression, of a Tet-on system to virus packaging, concentrating and purifying; incubating lentivirus with a typical swine trophoblastic cell line, changing fluid, adding puromycin for drug screening, diluting cells to single ones, and culturing to obtain the single-cell-source swine trophoblastic cell line with Cas9 expression under Tet-on regulation. The swine trophoblastic cell line is simple in operation and convenient to use and has a potential utilization value in researching of key functional genes of some trophoblast cells and screening of swine gene target spots. The cell line is expected to be a significant cell material for researching swine placenta development and applicable to correlation researches on swine early-stage placenta development and uterine pregnancy mechanisms and also has a high reference value in research of human early embryonic development.

The invention discloses a method for separating and purifying human embryo trophoblast and placental mesenchymal stem cells. The method includes: processing a placenta sample to obtain processed placenta tissues; digesting the tissues to obtain cell suspension; performing discontinuous Percoll density gradient separation to obtain trophoblast and placental mesenchymal stem cells; purifying the trophoblast; further separating and culturing the placental mesenchymal stem cells. The method has the advantages that HyQTase and DNAse I are used to jointly digest the tissues, and a large amount of trophoblast and placental mesenchymal stem cells with ideal cell purity and activity is obtained through the density gradient separation; digestion fragments, fibroblast and red blood cells are removed, the trophoblast and the placental mesenchymal stem cells are distinguished from each other, the purity of the trophoblast can reach 90%, the non-adherence upper layers of the placental mesenchymal stem cells are removed through a differential adhesion method, serum-free medium is added into a culture flask to continue the culture of the placental mesenchymal stem cells, and high purity of the placental mesenchymal stem cells is achieved.

The invention provides a multi-cell immune preparation for treating tumors and a preparation method of multi-cell immune preparation and relates to an immune preparation and a preparation method thereof. According to the multi-cell immune preparation and the preparation method, the problems that the treatment of existing immune cell preparations to the tumors is relatively limited, and the infusion safety cannot be guaranteed are solved. The multi-cell immune preparation comprises immune cells, human serum albumin and compound electrolyte. The preparation method comprises the following steps: (1) preparation of autologous plasma; (2) separation of peripheral blood mononuclear cells PBMC; (3) preparation of DC; (4) preparation of CIK; (5) preparation of NK cells; (6) preparation of CD3AK; (7) preparation of gamma delta T; (8) preparation of the multi-cell immune preparation. The multi-cell immune preparation has broad spectrum activity to the treatment of the tumors, the immune cells are derived from the body of a patient, and exogenous substances such as fetal calf serum, trophoblast cells and the like are not introduced, so that the infusion safety is guaranteed. The multi-cell immune preparation is applied to the field of tumor treatment.

The present invention relates to a method for separating free fetal cells from peripheral blood of a pregnant woman. The method utilizes the cell separation function and the specific affinity recognition principle of a microfluidic chip to realize high-efficiency and high-purity capture and separation of the free fetal cells (fetal nucleated red blood cells, trophoblast cells, and the like) in theperipheral blood of the pregnant woman. Within the microfluidic chip, a microarray structure with a specific geometric arrangement is designed, and the microarray structure is modified with moleculargroups having an affinity recognition function such as a nucleic acid aptamer, an antibody and a polypeptide. By adjusting parameters of the microarray structure, the collision efficiency of different sizes of fetal cells and microarrays can be controlled, and the capture and enrichment of the fetal cells of different sizes can be realized.

The invention discloses an umbilical cord blood Treg cell in-vitro amplification method based on trophoblastic cells and application. The specific technical method comprises the steps that firstly, umbilical cord Wharton's jelly mesenchymal stem cells are adopted as the trophoblastic cells to induce preliminary proliferation of Treg cells in umbilical cord blood mononuclear cells; then, pure Tregcells are obtained through magnetic bead sorting; and finally, the Treg cells are stimulated to be rapidly amplified by using optimized amplification factors. According to the amplification method, human AB plasma, IL-2, rapamycin, an RARA agonist and a DNA methyltransferase inhibitor are used as the optimized amplification factors, and a large number of umbilical cord blood Treg cells with high purity and high activity can be prepared within two weeks. Umbilical cord blood is used as a raw material for Treg cell amplification, batch preparation can be achieved, and Treg cell quality fluctuation caused by individual differences of samples can be reduced. The umbilical cord blood Treg cells have low immunogenicity and can be used as universal cells for clinical research, such as autoimmunediseases, graft-versus-host diseases and the like.

This invention discloses a method for separating and purifying skin caticular dry cells which adopts a self designed culture medium to suppress growth of fibrablasts and unform epithelioid cells grow around the tissue block to induce the cells to move out of the original micro-environment and gather again and grow on the new grown epithelioid cell layer. These clone grown cells are obviously different in morphology from its dependent trophocytes and can be selected with the help of dissection mirror and glass pins and nurtured for expansion independently. The acquired cells are tested by the generally acknowledged skin dry cells molecular label: ck19, integrin-beta1, P63, integrin-alpha 6 are positive which means the selected ones are purified skin dry cells.

The invention belongs to the field of genetic engineering, and in particular relates to application of PVT1 in preparation and diagnosis of preeclampsia and target drug therapy. The down-regulation of PVT1 in a placenta tissue of a preeclampsia pregnant woman is related to the generation and development of preeclampsia, and the low-expression-level PVT1 is closely related to the pathogenesis of preeclampsia. The proliferation, apoptosis, invasion, migration and the like of trophoblast cells of the preeclampsia pregnant woman are affected by changing the expression of PVT1, and the proliferation, invasion and migration of trophoblast cells can be promoted by enhancing the expression of PVT1.

The invention provides a method for diagnosing in a subject a condition requiring modulation of or involving trophoblast cell death, differentiation, invasion, and / or cell fusion and turnover, comprising detecting HIF Iα and the factors which modulate or are modulated by this protein, specifically TGFβ3, sFLT, VEGF, SMAD2, 3 and 7, MtdP, MtdL, MclI 1, MclIc, VHL, SiahI, Siah2, ENG, and PHD. The invention also provides a method for diagnosing or distinguishing in a subject a specific condition requiring modulation of or involving trophoblast cell death, differentiation, invasion, and / or cell fusion and turnover, in particular early onset severe preeclampsia (EPE), late onset preeclampsia (LPE) and mtre-uterme growth restriction (IUGR) comprising detecting HLF Iα and the factors which modulate or are modulated by this protein as defined above.

The invention discloses a mouse embryo stem cell culturing method and specific cultivating base, which cultures embryo stem cell on the trophoblast cell, wherein the trophoblast cell is epithelial cell or endothelium cell; the cultivating base consists of epithelial cell or endothelium cell and culturing liquid of embryo stem cell, which is high-sugar DMEM with 10-20% embryo cow serum or calf serum with 1. 5-3. 0*10-4mol / L Monothioglycerol, MTG.

The invention provides a preparation method of autologous natural killer cell proliferation and culture. The preparation method of the autologous natural killer cell proliferation and culture is characterized in that mononuclear cells of a cancer patient are amplified and activated to obtain natural killer cells after the mononuclear cells are transfected by 3- phosphoinositide-dependent protein kinase-1 and CD122 and under the effect of K562 cells which are expressed stably serve as trophoblast cells and interleukin 2, proliferation times of the natural killer cells are above 2500, phenotype of CD16+ / CD56+ is above 90%, and in-vivo and in-vitro tests show that the natural killer cells have an anti-tumor and have quite high killer toxicity. The invention also provides a kit which is used for autologous natural killer cell proliferation and culture and is obtained by using the preparation method. The kit has an efficient anti-tumor function.

The invention discloses a method for separating placental trophoblastic cells by using an immunomagnetic bead process. The method comprises the following steps: (1) preparing an immunomagnetic bead-coupled specific antibody by adding a coupling agent and an antibody capable of specifically capturing placental trophoblastic cells into immunomagnetic beads and carrying out incubation; (2) washing the antibody-coupled immunomagnetic beads obtained in the step (1) twice, and then preserving the antibody-coupled immunomagnetic beads in a preservation solution for subsequent use; (3) collecting a placental trophoblast sample and preparing a sample cell suspension by using an enzymolysis process; (4) washing the sample cell suspension obtained in the step (3) twice; (5) adding the antibody-coupled immunomagnetic beads treated in the step (2) and carrying out incubation; and (6) washing the immunomagnetic beads having undergone incubation in the step (5) twice and discarding a supernatant so as to obtain purified placental trophoblast cells. According to the invention, immunomagnetic bead cell sorting technology is employed, and the antibody-coupled immunomagnetic beads are used for specific capture of placental trophoblast cells, so trophoblastic cells can be easily and rapidly separated via a few simple steps, and automation can be easily realized.

The invention provides an NK cell culture system and an application thereof, and belongs to the technical field of immune cell therapy. The NK cell reinfusion solution is prepared through the steps of peripheral blood mononuclear cell separation, cell activation, proliferation and the like by utilizing the NK cell culture system, exogenous animal serum does not need to be added, exogenous trophoblast cells do not need to be adopted either, and the preparation method is safer, easy to operate, low in cost, and suitable for clinical-grade production of the NK cells. The tumor killing experiments show that the NK cell culture system can relieve NK cell inhibition to a greater extent, activate NK cells and improve the tumor killing activity of the NK cells.

The invention provides an autologous dendritic cell activated tumor-infiltrating T-lymphocyte preparation method. The method has the following characteristics that: a tumor-infiltrating T-lymphocyte from a cancer patient is activated and amplified in a large quantity by using a dendritic cell loaded by an autologous tumor antigen as a trophoblast cell and interleukin-2, and has a highly active anti-tumor cell toxic response. The invention further provides an autologous tumor-infiltrating T-lymphocyte kit prepared according to the method, and the kit has an efficient anti-tumor function.

The invention provides a method for separating trophoblastic cells. The method comprises the following steps: obtaining specific antigens expressed on surfaces of various types of trophoblastic cellsaccording to searched documents, and screening the obtained specific antigens; confirming expression of the antigens in the trophoblastic cells through immunohistochemistry, and determining an antibody combination system through immunofluorescence; adding a couplant and an antibody combination into immunizing magnetic beads, and carrying out coupling; collecting a trophoblast sample from a placenta, preparing a sample cell suspension from the sample, and adding the immunizing magnetic beads with a specific antibody for incubation; and washing the immunizing magnetic beads after the incubationstep is completed, thereby obtaining separated purified trophoblastic cells of the placenta. Compared with traditional amniocentesis and chorionic villus sampling, the method has a noninvasive advantage, the material drawing time is relatively early, the risk of infection and abortion is low, and detection results have similar reliability.