Figure 5. In vitro and in vivo differentiation of H9 hES cells cultured on bFGF-hFLSCs for 10 passages (about 70 days).

(A): Floating culture of H9 hES cells at day 6. Bar: 100 µm. (B–D): Images of differentiated cells at day 10 (B), epithelial cells (C), fibroblast like cells (D). Bars: 100 µm. (E): RT-PCR analysis of various differentiation markers for the three germ layers and extraembryonic trophoblast. (F): Immunocytochemistry of AFP, a-SMA, and βIII-TUBULIN. Nuclei were stained with DAPI (blue). Bars: 100 µm. (G): Teratoma formation. bFGF-hFLSCs-cultured H9 cells for 10 passages (about 70 days) form teratoma generating structures representative of three germ layers. Bars: 100 µm.