Fig 3.

Phenotypic characterization of FOXA2 expressing neural progenitor cells from RA-treated human ES/iPS cells. (A–L) Immunocytochemistry at DIV19 revealed that human ES cell-derived neural progenitor cells formed rosette-like structures and coexpressed OTX2 (red) but not FOXA2 (green) in response to Fgf8b with (A, E, I) or without mouse Shh (D, H, L). In contrast, isolated neural rosette-like structures were observed in cultures exposed to SHH-C24II and FGF8a (B, F, J). These rosette-like structures coexpressed FOXA2 and OTX2 (B). In cultures exposed to SHH-C24II and Fgf8b, very few rosette-like structures were observed and cells coexpressed FOXA2 and OTX2 (C, G, K). (M) Human ES cells exposed to 500 ng/ml SHH and FGF8a or Fgf8b differentiated into FOXA2⁺ cells (green) that expressed human-specific nestin (red). (N) In similar culture conditions, FOXA2⁺ cells (green) coexpressed 3CB2 (red). (O) In parallel cultures, all human iPS cell lines differentiated into FOXA2⁺ (red)/β-tubulin^Low (green) neural progenitor cells. Scale bar A–L, O = 50 µm, M = 20 µm, N = 10 µm.