Effects of ethanol on the differentiation of hESC towards hepatocytes.

<p>(A) Schematic illustration of the differentiation protocol and the time frame of ethanol treatment. (B) Immunostaining of albumin in the hESC-derived hepatocytes (hEH) at day 24 exposed to the indicated doses of ethanol (x100). (C) Cell viability was assessed at day 24 by MTT assay. (D, E) qPCR was performed to determine the expression of liver genes, ALB and ASGPR1, C/EBPα, and HNF1α at day 24 exposed to the indicated doses of ethanol. (F) Secreted ALB levels in the culture media by hEH were assessed by ELISA at day 24 with or without ethanol treatment. (G) Dynamic expression changes of metabolizing enzymes, CYP3A4, CYP7A1, and UTG1A6 in hEH were evaluated by qPCR at different time points after differentiation in the presence or absence of ethanol at 100 mM. (*p<0.05 vs. no EtOH; #p<0.005 vs. no EtOH).