Fig. 2

Screening for induction protocol to generate iHeps from ADSCs. a Sequential morphological changes from ADSCs to iHeps. I Cell morphology before hepatic lineage induction. II Change of cell morphology with a short spindle shape after culture in endodermal induction medium. III At day 4, the cell morphology had become polygonal in shape. IV Morphology of the mature iHeps. b Immunostaining revealed the vast majority of induced cells were positive for the DE markers Sox17 and FoxA2 at day 1 (upper right, ADSCs). c Differentiation efficiency measured by flow cytometry analysis marked by ALB, AAT, and ASGPR1. n = 3. d Gradual changes of hepatocytic gene expression in iHeps during hepatic lineage induction. The expression levels of the indicated genes were analyzed by quantitative PCR. Data are normalized to iHeps at day 4. AAT alpha-1-antitrypsin, ALB albumin, ASGPR asialoglycoprotein receptor, CK18 cytokeratin 18, d day, GJB1 gap junction protein beta-1, HNF4α hepatocyte nuclear factor alpha-4, rHep primary rat hepatocyte, TAT tyrosine transaminase, TTR transthyretin, AFP alpha fetal protein. Scale bar: 100 μm (a,) 50 μm (b). Data presented as mean ± standard deviation