A culture method for maintaining the self-renewal state of human embryonic stem cells
A technology for human embryonic stem cells and embryonic stem cells, applied in the field of embryonic stem cell culture conditions, can solve problems such as unfavorable exploration, unstable state of human embryonic stem cells, and inconvenient passage operations.
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[0070] (1) HES2 human embryonic stem cells (Human Embryonic Stem Cells, hESCs) were provided by Professor Qilong Ying from the University of Southern California;
[0071] (2) Passage and culture of HES2 human embryonic stem cells under the condition of Activin A+BFGF+IWR1:
[0072] a. Take a 6-well cell culture plate, wrap each well with 2ml DMEM+10μl Matrigel, and place at 37°C, 5% CO 2 Concentration in the cell culture box, coated for 4 hours;
[0073] b. Take the human embryonic stem cells grown to 70-80% density, discard the culture medium, and wash the cells once with PBS to remove the residual culture medium on the cell surface;
[0074] c. Add 1ml of CTK to digest the embryonic stem cells. After about 7 minutes, the edge of the cells floats. Discard the CTK, add 2ml of conventional cell culture medium containing 10% FBS, blow and blow the cells from the culture plate with a pipette, and then transfer into a 15ml sterile centrifuge tube;
[0075] d. After centrifugati...
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