Culture method related to differentiation of pluripotent stem cells into blood cells
A cell culture and blood cell technology, applied in the direction of cell culture active agents, artificially induced pluripotent cells, biochemical equipment and methods, etc., can solve the problems that there is no precedent, and achieve the effect of high blood differentiation ability
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[0096]
[0097] 1. Cells and Reagents
[0098] Human iPS cells used TkCB7-2, TkCB7-4, and TkCB8-3 established by introducing Yamanaka 4 factors (OCT3 / 4, KLF4, SOX2, and c-MYC). The maintenance culture of iPS cells was performed by adding 0.1 mM non-essential amino acids (Invitrogen), 2 mM L-glutamine (Invitrogen), 20% knockout serum substitute ( Invitrogen), 0.1 mM 2-mercaptoethanol (SIGMA), and 5 ng / mL basic fibroblast growth factor (bFGF; Upstate). iPS cells were cultured on mouse fetal fibroblasts (MEFs) irradiated with radiation (50Gy), and passaged every 3 days to maintain an undifferentiated state. In addition, mouse C3H10T1 / 2 cells used as feeder cells were cultured by adding 10% fetal bovine serum (FBS) and 2 mM L-glutamine to Eagle basal medium (Invitrogen).
[0099] The differentiation medium was Iscove's modified Dulbecco's medium (SIGMA) with 10 μg / mL human insulin, 5.5 μg / mL human transferrin, 5 ng / mL sodium selenite (SIGMA), 2 mM L-glutamine, 0.45 mM α - Mon...
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