Differentiation of human umbilical cord mesenchymal stem cells into functional hepatocyte induced by liver homogenate supernatants in vitro

Abstract

Abstract: Objective To investigate functional characteristics of hepatocytes that differentiating from human umbilical cord mesenchymal stem cells (hUCMSCs) induced by liver homogenate supernatants (LHS). Methods The 3rd Passage of hUCMSCs were selected and divided into control group and LHS group. At three time points (3d, 5d, 7d) after being induced, the immunofluorescence-cytochemistry staining of alpha-fetoprotein (AFP), cytokeratin 18 (CK18), tryptophan hydroxylase 2 (TPH2), cytochrome P450 3A4 (CYP3A4), albumin (Alb) and hepatocyte growth factor (HGF) were performed, and the CYP3A enzyme activity and levels of HGF, Alb and BUN were detected. Results The cells had expressed liver specific protein at the third day after LHS inducing, including AFP, CK18, TPH2, CYP3A4, Alb and HGF. In LHS group, the metabolism yield of midazolam, the specific metabolic substrates of enzyme CYP3A was increased significantly than that in control group (30.14±1.19, 50.20±6.24 and 120.85±15.52 ng/mg versus 0.026 to 0.030 ng/mg, P<0.01). Compared with control group, the secretion levels of HGF and Alb in LHS group were increased significantly at 3 time points. Additionally, the urea production of cells in LHS induction group increased 14 times more than that in control group (P<0.01). Conclusion The study has demonstrated that hUCMSCs could differentiate into hepatocytes with mature functions in vitro after LHS induction.

Key words: Human umbilical cord mesenchymal stem cells, Inducting and differentiation, Liver homogenate supernatants, Hepatocyte

XUE Gai, HAN Hua, CHEN Ze-sha, WU Hong-hai, LIU Jian-fang, HOU Yan-ning. Differentiation of human umbilical cord mesenchymal stem cells into functional hepatocyte induced by liver homogenate supernatants in vitro[J]. Chinese Hepatolgy, 2016, 21(9): 733-737.

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