Hepatocyte growth factor and epidermal growth factor induce mesenchymal stem cells isolated from patients with hepatocirrhosis into hepatocyte-like cells in vitro / 第三军医大学学报

ABSTRACT

Objective To explore the possibility of differentiation of mesenchymal stem cells isolated from patients with hepatocirrhosis into hepatocyte-like cells induced by EGF and HGF and to lay basis for transplanted autologous bone marrow MSCs in treatment of liver disease at terminal stage.Methods Bone marrow cells were obtained from volunteers with liver cirrhosis.MSCs were separated by density gradient centrifugation and were cultured through adhere culture.MSCs were cultured in DMEM medium with HGF,EGF,HGF+EGF or no growth factor.The phenotypes of MSCs were identified by flow cytometry,immunohistochemistry,and Albumin levels in culture supernatants were determined by ELISA.Results Growth and division of adherent cells obtained from the patients with hepatocirrhosis were good and the phenotypes of MSCs were CD29 positive and CD34 negative.The shape of MSCs changed from long fusiform to polygonal or round on 21th-28th days in grow factor induced groups.Immunocytochemical analysis for CK18 and AFP showed positive staining reaction for AFP on 7th day,for CK18 on 21st and 28th day in grow factor induced groups with MSCs-induced Alb production increasing in a time-dependent manner.No markers of hepatocyte linear cells were detected in no growth factor induction group.Conclusion Both HGF and EGF can induce mesenchymal stem cells to differentiate hepatocyte-like cells alone or coordinately.