84 results about "Feeder Layer Cells" patented technology

The invention provides a culture system which contains defined components and efficiently acquires induced pluripotent stem cells (iPS cells) from somatic cells. The culture medium additive provided by the invention contains vitamin C, vitamin B12, insulin, glycogen synthase kinase-3 inhibitor, receptor tyrosine kinase and antioxidant. The culture medium additive provided by the invention can also contain a substituted serum cell growth promoter. The invention also provides a complete medium acquired by inducing pluripotent stem cells, which is prepared from one or more of basic culture medium, serum and substituted serum additive and the culture medium additive. The culture system provided by the invention does not have serum, has no animal source pollution, contains defined chemical components, efficiently acquires iPS cells, can maintain cell growth and proliferation in the process of conversion from somatic cells to iPS cells under the condition that feeder cells do not exist, simultaneously accelerates the induction course of the iPS cells obviously, and greatly improves the efficiency of inducing the somatic cells into the iPS cells.

Undifferentiated primordial stem cells are manipulated to permit their long term growth in defined media lacking serum and feeder layer cells by shifting the apoptotic balance of the cells, through increasing the activity of Bcl-2 family anti-apoptotic proteins or decreasing the activity of Bcl-2 family pro-apoptotic proteins. In some embodiments of the invention, the Bcl family protein is Bcl-2. The ES cells sustain the characteristics of undifferentiated, pluripotent stem cells during long-term serum- and feeder layer cell-free conditions, including the ability to be expanded in vitro, but maintain their potential to differentiate into mature cell types.

Dopamine-producing neural precursor cells are produced by: producing a cell population containing Corin and / or Lrtm1 positive cells via steps (1) and (2) as will be shown below; collecting the Corin and / or Lrtm1 positive cells from the cell population thus obtained by using a substance that binds to Corin and / or a substance that binds to Lrtm1; and suspension-culturing the Corin and / or Lrtm1 positive cells in a liquid culture medium that contains one or more neurotrophic factors: (1) a step for adhesion-culturing pluripotent stem cells in the absence of any feeder cells in an undifferentiatedstate-maintaining culture medium that contains a sonic hedgehog (SHH) signal stimulant and an undifferentiated state-maintaining factor in the presence of an extracellular matrix; and (2) a step for culturing the cell population obtained in step (1) in a liquid culture medium that contains one or more differentiation inducing factors.

The invention relates to a method for preparing breast cancer bone metastasis organs, which comprises the following steps: mixing breast cancer bone metastasis cells, feeder layer cells, a temperature-sensitive hydrogel and a first culture medium, and culturing to obtain breast cancer bone metastasis organs; wherein the feeder layer cells comprise bone marrow mesenchymal stem cells without a proliferation function. By means of the technical scheme, the bone marrow mesenchymal stem cells without the proliferation function are used as feeder layer cells for breast cancer bone metastasis organ culture, and the success rate of breast cancer bone metastasis organ culture through the method is high.

The invention relates to a method for directionally inducing and differentiating induced pluripotent stem cells into vascular endothelium-like cells. According to the method, a culture system without feeder cells is adopted, the vascular endothelium-like cells can be obtained only through expressing FLI1 related genes and activating a protein kinase C pathway, and a test shows that the purity is relatively high. Through the adoption of the method for directionally inducing and differentiating induced pluripotent stem cells into vascular endothelium-like cells, the induction period is short, the efficiency is high, the obtained vascular endothelium-like cells are stable in performance, and the problem of matrix cell pollution is avoided.

The invention provides a stem cell culture perfusion device which comprises a perfusion bottle, a feeder layer cell culture unit, a cell perfusion tank and a waste liquid bottle, wherein the opening of the to-be-cultured cell perfusion bottle is provided with a first filter head and an input tube; the input tube is provided with a first controller capable of displaying pH value and controlling temperature; the tail end of the input tube is immersed in the feeder layer cell culture unit; a first connecting tube, which is connected through the feeder layer cell culture unit of which the substrate is provided with a semi-permeable membrane, is provided with a second controller and a first peristaltic pump; both ends of the cell perfusion tank are respectively connected with a second connecting tube and a third connecting tube; the other end of the second connecting tube is connected with the first peristaltic pump; the other end of the third connecting tube is connected with the a second peristaltic pump; the opening of the waste liquid bottle is provided with an output tube and a second filter head; and the other end of the output tube is connected with the second peristaltic pump. Under the actions of the first controller and second controller, the temperature and pH value variations can be observed, the rate of the peristaltic pumps can be controlled, and error values caused by manual observation and recording can be avoided; and the equipment output of the cell perfusion tank can enhance the stem cell culture rate.

The invention relates to the field of cell biology technologies and application, and particularly relates to a stomach cancer tumor primary organ culture method. The method comprises the steps of obtaining feeder layer cells and stomach cancer tumor primary cells by adopting an enzyme digestion method and cell culture method; adding magnetic suspension particles to the feeder layer cells and the stomach cancer tumor primary cells respectively, and obtaining the magnetic particle containing feeder layer cells and stomach cancer tumor primary cells by incubation-digestion; inoculating the magnetic particle containing feeder layer cells and stomach cancer tumor primary cells to a stomach cancer tumor primary organ culture medium, executing culture under the magnetic driving conditions to obtain stomach cancer tumor primary organs. By adopting a magnetic suspension culture system, the feeder layer cells and stomach cancer tumor primary cells are co-cultured, the mutual effect of tumor microenvironment and the cells is better simulated, and the new in-vitro efficacy testing method is provided for the drug research and development industry.

The invention discloses a method for preparing feeder layer cells. The method provided by the invention comprises the following steps: (1) treating the isolated fibroblasts with 10 μg / ml mitomycin C, collecting the culture supernatant and adherent cells respectively; (2) treating the isolated fibroblasts The cells are treated with the culture supernatant collected in the previous step, and the adherent cells are collected; between the step (1) and the step (2), the following steps of 0 to 2 times are also included: the isolated fibroblast The cells are treated with the culture supernatant collected in the previous step, and the culture supernatant and adherent cells are collected respectively; the adherent cells obtained in each of the above steps are feeder layer cells. The method for preparing feeder layer cells provided by the invention has simple flow, economical and reliable effect. Human embryonic stem cell culture experiments show that the feeder layer cells prepared by the method can be passed on for a long time, and can effectively support the growth of human embryonic stem cells and keep them in an undifferentiated state.

The invention discloses a fetus liver matrix cell strain for expressing alkaline fibroblastic growth factors. The fetus liver matrix cell strain of the invention is a recombinant fetus liver matrix cell strain carried with alkaline fibroblastic growth factor genes. By experimental verification, recombinant fetus liver matrix cells of the invention can highly efficiently produce secretory alkaline fibroblastic growth factors, and embryonic stem cell proliferation of human beings and embryonic stem cell drying can be maintained under the condition of no exogenous alkaline fibroblastic growth factors by taking the cells as feeder layer cells for in vitro culture of embryonic stem cells of human beings. The recombinant fetus liver matrix cells of the invention and a construction method thereof provide appropriate microenvironment for researches in the aspects of construction of a low-cost (reduction of introduction of exogenous alkaline fibroblastic growth factors), safe and highly efficient (capable of replacing odontogenic zooblast as the feeder layer cells) in vitro culture system of the embryonic stem cells of human beings and development and differentiation of hematopoietic cells, and the invention has wide application prospect.

The invention provides new application of rabbit fetal fibroblast, and particularly relates to a method for preparing a ZEN (Zearalenone) monoclonal antibody by utilizing the rabbit fetal fibroblast.The method is characterized in that mouse hybridoma is cultured by using rabbit fetal fibroblast as feeder cells of murine hybridoma, the murine feeder cells can be completely replaced, growth of monoclonal hybridoma can be promoted, and a monoclonal antibody with high valence, good stability and strong specificity can be generated; the hybridoma bred by different animal cells as the feeder cellsis used for preparing an antibody, and very high valence and very high specificity are obtained; meanwhile, the rabbit fetal fibroblast can be revived by many times for use after being cryopreserved,animal tissue isolating culture is not required to be carried out every time. The method disclosed by the invention is simple, the cost is low, the monoclonal antibody which is used for fast, sensitive, economic and efficient ZEN immunological detection can be prepared, and a wide application prospect and huge market value are obtained.