Fig. 5

Dlk1⁻ cells appeared to be cholangiocyte progenitor-like cells. Dlk1⁺ cells and Dlk1⁻ cells derived from Dlk1⁺ cells were sorted using FACS. a, b Dlk1⁺ cells and Dlk1⁻ cells were induced for hepatic differentiation with Matrigel and OSM. Phase-contrast images presented the morphologies of the cultured Dlk1⁺ cells and Dlk1⁻ cells on Matrigel after 8 days (a). Arrows indicate differentiated cells. b Cells engaged in hepatic differentiation for the indicated time were collected, and the expression of the hepatic maturation markers ALB, G6P, and TAT was determined on day 2 and TO expression was measured on day 8 via quantitative real-time PCR. c, d Dlk1⁺cells and Dlk1⁻ cells were induced for cholangiocytic differentiation in collagen gel-embedded cultures. Phase-contrast images show representative views of branching structures derived from Dlk1⁻ cells compared with Dlk1⁺ cells (c). Arrows indicate bile duct-like branching structures. Cells engaged in cholangiocytic differentiation for the indicated times were collected, and the expression of the cholangiocyte marker CK19 was determined with quantitative real-time PCR (d). Representative images from one of three experiments are shown, and the data are shown as the mean ± SEM of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001 (Mann-Whitney test and two-way ANOVA with post Bonferroni’s multiple comparisons test)