170 results about "Trophoblast cell" patented technology

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 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 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 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 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 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 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 belongs to the technical field of primary cell culture, and especially relates to a B lymphocyte in vitro culture system and applications thereof. The culture system utilizes a T lymphocyte line capable of secreting interleukin molecules such as IL-2 as trophoblastic cells so as to provide micro interleukin molecules required by the growing development of B lymphocyte; and trimer CD40L molecules expressed in vitro through a recombinant manner and having high biological activity are added as important molecules so as to further stimulate the growth, reproduction and development ofthe B lymphocyte. The system can also perform gamma ray irradiation treatment on the trophoblastic cells, so that the pre-treated trophoblastic cells can lose the ability of cell reproduction but still has the ability of protein synthesis and secretion within 7-14 days. The provided system can effectively culture the B lymphocyte derived from rabbits and alpacas in vitro, and can utilize the method to develop corresponding monoclonal antibodies.