3.4. Endothelial Cell Induction (Days 4 to 6)

Starting day 1, the cells are always incubated in a normoxic tissue culture incubator (20% O₂, 5% CO₂, 37°C).

(a) Human iPSCs plated as a monolayer on day 0 of differentiation undergo morphological changes while continue to proliferate. Between day 1 and 4 of differentiation, the density of culture increases, forming multilayers of cells. On day 5, the cells acquire an endothelial cell-like phenotype and can be harvested for magnetic automated cell sorting (MACS) on day 7. (b) Following MACS enrichment, the CD31⁺ iPSC-ECs can be expanded on collagen-coated plates for multiple passages. Scale bar, 50 μm