Differentiation of hPSCs into definitive endoderm cells on various substrates.

<p>(A) Most cells expressed the pluripotency marker POU5F1 on the hE-cad-Fc-coated surface, but almost all cells differentiated into definitive endoderm cells expressing GATA4 on vitronectin- and Matrigel-coated surfaces. (B) The vitronectin-coated surface supported differentiation of K3 cells into ALB⁺ hepatocyte-like cells at day 20 of differentiation. (C) Differentiating cells detached from the vitronectin-coated surface at day 10, although these cells expressed HNF4A. Scale bar indicates 100 μm. (D) Quantitative PCR analyses of cadherin and MMP gene expression during differentiation of human iPS cells. 201B6 cells were differentiated into hepatocyte-like cells on an R-Fc-coated surface, and total RNA was isolated at days 0, 5, 10, 15, and 20 of differentiation. Data are the means ± SD of three independent differentiation experiments. *p < 0.001, **p < 0.005 vs. day 0 (d0) sample.