Fig. 6

Differentiation of proliferative HBs into functional hepatocytes. A The morphology of small-molecule protocol-derived HLCs. HLCs exhibited polygonal shapes and distinct round nuclei; even some had two nuclei. Scale bar 100 μm. B Quantitative RT-PCR analyzed the genes expression levels of ALB, the CYP and urea cycle enzymes. Gene expression was normalized to HBs. Data are presented as mean ± SEM, n = 3. * P < 0.05, ** P < 0.01. C Immunostaining analyses of mature hepatocyte markers (ALB, A1AT, CYP3A4, and CYP2C9) on small-molecule protocol-derived HLCs. Scale bars 100 μm. D CYP450 activity assay of HLCs. Data are presented as mean ± SEM, n = 3. * P < 0.05, ** P < 0.01. E Urea secretion of HLCs was analyzed. Data are presented as mean ± SEM, n = 3. * P < 0.05, ** P < 0.01. F PAS staining on HLCs. Scale bar represents 200 μm. G ICG uptake analyses in HLCs. Scale bar represents 200 μm. H The HLCs were stained by Alexa-Flour 488-ac-LDL. Scale bar 200 μM