Fig. 2

Direct reprogramming of MSCs to d-iHEPs using dox-inducible Foxa2. Schematic diagram depicting the procedure and time-course for the direct conversion of MSCs into d-iHEPs (a). Colonies of epithelial-like d-iHEPs (red dashed circles) with GFP expression are observed after 15 days of iHEP medium exposure (b). Purified d-iHEPs at passage 6 expressing the hepatic markers albumin (ALB) and CK18 (green) by immunofluorescence. Nuclei are visualized in blue by DAPI staining (c). The increased presence of fibroblast-like cells after continuous passaging in the absence of doxycycline (d). RT-qPCR showing decreased levels of Foxa2 mRNA in d-iHEPs when compared to iHEPs (e). Validation of Tet-on Foxa2 system by analysis of Foxa2 expression in iHEPs and d-iHEPs in the presence or absence of doxycycline, by RT-PCR (f). ***p < 0.001. Scale bars 50 μm