Fig. 3
From: Transplantation of hESC-derived hepatocytes protects mice from liver injury
Differentiation of VAL9-hepatoblasts into hepatocytes. a Protocol and phase contrast images of hepatocyte differentiation. Hepatic progenitors were passaged at day 11 on collagen 1-coated wells and grown for 2 days in HamF12/Williams (HPM), 20 ng/ml hepatocyte growth factor (HGF), then for 2 days in HPM, 20 ng/ml HGF and 20 ng/ml epidermal growth factor (EGF). From day 16 to day 18 of differentiation cells were grown in a mixture of HPM/hepatocyte culture medium (HCM), HGF 10 ng/ml and oncostatin 10 ng/ml. Hepatocytes were generated after 10–12 additional days in HCM, 10 ng/ml HGF. b Hepatic morphology of VAL9-HEP during the differentiation protocol (11 to 30 days). c Representative field of immunostaining of VAL9-HEP. Cells express hepatocyte nuclear factor (HNF)Aα, alpha-1-antitrypsin (A1AT), albumin (ALB; red) and HNF3β (green), ALB (red) and cytochrome P450 3A4 (CYP3A4), alpha foetoprotein (AFP), claudin (CLDN1), scavenger receptor class B member 1 (SRB1) and cluster of differentiation (CD)81. d Representative fluorescence-activated cell sorting analysis of VAL9-HEP showing 85 % of asialoglycoprotein receptor (ASGR) a marker specific for differentiated hepatocytes. e Quantitative RT-PCR analysis at days 0, 4, 11 and 30 of differentiation. Data are represented as the percentage of expression in VAL9-HEP. f Representative transcript levels of hepatic cell markers on days 5, 11 and 25 of differentiation