Fig. 1.

Generation and phenotypic characterization of iHEPs. Schematic diagram depicting the procedure and time-course for the direct conversion of MSCs into iHEPs (a). Phase-contrast and fluorescence microscopy images representing MSCs’ morphology before hepatic reprogramming and, at reprogramming day 15, the appearance of colonies of epithelial-like cells with GFP expression (b). Phase-contrast image of a purified iHEP culture and respective growth curve at P8 (c). Characterization of iHEPs by immunofluorescence showing expression of E-cadherin and FoxA2 (green) and albumin and CK18 (red). Nuclei are visualized in blue by DAPI staining. Wild-type MSCs were used as controls (d). RT-qPCR analysis of MSCs, iHEPs, fetal hepatoblasts (E13.5), and adult primary hepatocytes for hepatic markers (e). Data are shown as mean ± SEM of three independent samples for each group. *p < 0.05; **p < 0.01; ***p < 0.001. Scale bar 100 μm