Figure 3.

Hepatocyte functional assay after hepatocyte and nonparenchymal cell codifferentiation. (a) Quantitative PCR analysis showed the expression levels of mature hepatocyte markers.  ^∗p < 0.05,  ^∗∗p < 0.01,  ^∗∗∗p < 0.001, and n = 3. (b, c) Immunofluorescence showed the coexpression of mature hepatocyte markers in day 25 cells. Scale bars, 50 μm.  ^∗p < 0.05,  ^∗∗p < 0.01,  ^∗∗∗p < 0.001, and n = 3. (d, e) Hepatocytes performed PAS staining (top), ORO staining (middle), and ICG uptake/release analysis (bottom). Scale bars, 25 μm.  ^∗p < 0.05,  ^∗∗p < 0.01,  ^∗∗∗p < 0.001, and n = 3. (f, g) Phase-contrast images of senescence β-Gal staining in hepatic cells during days 35-45 treated with Wnt3a or BMP4. Scale bars, 25 μm.  ^∗p < 0.05,  ^∗∗p < 0.01,  ^∗∗∗p < 0.001, and n = 3. (h) Production of ALB and AAT in day 35 hepatocytes. Primary hepatocytes were used as positive controls; hiPSCs were used as negative control.  ^∗p < 0.05,  ^∗∗p < 0.01,  ^∗∗∗p < 0.001, and n = 3.