Generation of Hepatocytes and Nonparenchymal Cell Codifferentiation System from Human-Induced Pluripotent Stem Cells

Figure 5

Optimization of the BMP4 differentiation protocol significantly promotes hepatocyte function. (a) q-PCR analysis showed the expression levels of mature hepatocyte markers. , , , and . (b–d) Immunofluorescence showed that the cells coexpressed hepatocyte markers induced by BMP4 or BMP4-optimized protocols. Scale bars, 50 μm. , , , and . (e, f) Hepatocytes performed PAS (top), ORO (middle), and ICG (bottom). Scale bars, 50 μm. , , , and . (g, h) Phase-contrast images of senescence β-Gal staining in hepatocytes during days 30-60 induced by original or BMP4-optimized medium. Scale bars, 25 μm. , , , and . (i) ORO staining treated by BMP4 optimized medium in day 65. Scale bars, 50 μm. (j) Immunofluorescence showed that hepatocytes coexpressed ALB and CYP3A4 in day 65. Scale bars, 25 μm. (k) Production of ALB and AAT in day 35 hepatocytes. Primary hepatocytes were used as positive controls; hiPSCs were used as negative control. , , , and .

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