Fig. 4

Generation of functional HLCs by two-step chemical strategy. a Schematic of the two-step chemical strategy to generate functional HLCs. b Quantitative RT-PCR analyzed the genes expression levels of the urea cycle and CYP 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, CYP3A4, and CYP2C9) on HLCs derived from two methods. Scale bars 100 μm. d Efficiency of hepatocyte markers expression, determined by counting positive cells. Efficiencies are presented as the percentage of positive cells plus or minus the SD of all fields counted. e CYP450 activity assay of different origins of hepatocytes. Data are presented as mean ± SEM, n = 3. *P < 0.05, **P < 0.01. f Urea secretion among different origins of hepatocytes were analyzed. Data are presented as mean ± SEM, n = 3. *P < 0.05, **P < 0.01. g PAS staining on different origins of hepatocytes. Scale bar represents 100 μm. h ICG uptake analyses in different origins of hepatocytes. Scale bar represents 100 μm