Abstract：Objective: Exploring the optimal conditions of human induced pluripotent stem cells(hiPSCs) committed differentiation into hepatocytes under twodimensional condition in vitro, and differentiation under threedimensional condition. Methods: The hepatocyteinduced differentiation medium was screened and optimized, and the optimal scheme was obtained by measuring the expression of albumin and AFP under four different differentiation medium.The hiPSCs were cultured under twodimensional condition.The expressions of liver marker protein were detected by immunofluorescence and Westernblotting.ELISA was used to investigate the function of hepatocytes.The hiPSCs were cultured under threedimensional condition.Cell morphological changes were observed by HE staining, and compared the function of hepatocytes by ELISA under two and threedimensional condition. Results: After 23 days of differentiation under two dimensional condition, the cells observed in inverted microscope showed typical hepatocytelike morphology, and positive expression of AFP, albumin, CK8, CK18 and SOX17 by immunofluorescence and Westernblotting, which was similar to that of embryonic stemderived hepatocytes(hESCsHEPs).The result of ELISA showed that cells differentiated from hiPSCs had normal function, indicating that hiPSCs had differentiated into hepatocytes. The results of HE staining showed that cells differentiated from hiPSCs had obvious dual nuclei under threedimensional condition, and it was the morphology of typical hepatocyte.The results of ELISA showed that the function of cells were not abnormal. Conclusion: In the two and threedimensional conditions,hiPSCs can be induced to differentiate into hepatocytes, expressing liverrelated proteins, and had normal function.
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2018, Vol. 28 
