Research Article

Fine Tuning of Hepatocyte Differentiation from Human Embryonic Stem Cells: Growth Factor vs. Small Molecule-Based Approaches

Figure 4

Hepatocyte differentiation of growth factors/NaB/DMSO-derived DE cells (5 d). (a, c, & e) The DE cells derived from growth factors/NaB/DMSO for 5 days were cultured in hepatic progenitor media without extra HGF added for 7 days and then cultured in L-15 hepatocyte maturation media (a), Touboul’s maturation media (b), or with HGF in hepatic progenitor media and Carpentier’s maturation media (c) for another 7 days. The cells were fixed on day 13 and day 20 of differentiation and photographed for phase images. The cells were then stained and imaged by a fluorescence microscope using antibodies against AFP, HNF4α, and ALB. DAPI represents nuclear staining. . (b, d, & f) The cells as explained above in (a), (c), & (e), respectively, were analyzed for mRNA expression by RT-qPCR for the indicated genes using gene-specific primers. The bars represent normalized (18S rRNA) fold mRNA expression. The data are represented as . (g) Comparison of mRNA expression analysis by RT-qPCR for the indicated genes from RNA samples as described for (a), (c), & (e), respectively.