Fig. 2

Three cultured liver progenitor cells are functionally distinct in vitro & in vivo. a Heat map showing known hepatocyte and biliary gene expression in three cultured LPCs, HC, and BEC. b Quantification of HNF1A and EpCAM positive cells among three cultured LPCs, assessed by flow cytometry. Red peaks represent staining samples and blue peaks represent the isotype control. c Representative images of organoids formed by three cultured LPCs. Scale bars, 100 μm. d Normalized expression levels of bile duct markers genes in LPCs derived organoids, analyzed by RT-q-PCR. e Representative immunofluorescence staining of EpCAM and CK19 and uptake of Rhodamine 123 dye in organoids formed by BecLPCs at day 10. Scale bars, 100 μm. f Representative images of spheroids formed by three cultured LPCs. Scale bars, 100 μm. g Normalized expression levels of hepatic and progenitor marker genes in LPCs derived organoids, analyzed by RT-q-PCR. h Representative PAS staining, and immunohistochemical staining for CYP3a4 and ALB of 3D spheroid formed by three cultured LPCs, respectively. Scale bars, 100 μm. i Schematic overview of the experimental design. The repopulated three cultured LPCs were analyzed by immunofluorescence staining for FAH expression, after transplantation of LPCs into Fah^−/− mice at day 30, as described in the Method. Scale bars, 600 μm. j The maturation of repopulated HepLPCs was analyzed by co-immunofluorescence staining for FAH and ALB expression at day 30 after transplantation of LPCs into Fah-/- mice, as described in the Method. Scale bars, 100 μm. Resident LPCs-derived LPCs, reLPCs; Hepatocytes-derived LPCs, HepLPCs; BECs-derived LPCs, BecLPCs. For panels d and g, results are shown as the mean ± S.D. of three independent experiments. *P < 0.05; **P < 0.005; ***P < 0.001.