Myofriboblast Promotes the Proliferation and Differentiation of Mouse Embryonic Hepatic Stem Cells

Abstract: Microenvironment is very important to proliferation and differentiation of transplanted cells.As a crucial composition of the microenvironment in tumor， myofibroblast influence the development of tumor cells deeply. This study focuses on the effect of fibroblast and myofibroblast on the proliferation of embryonic hepatic stem cells (EHSCs) HP14.5. In this research， two sorts of conditioned medium were harvested from the supernate of fibroblast (CMFb) and the supernate of myofibroblast (CMAFb) which came from the Hepal-6 induced differentiation of fibroblast in vitro. The EHSCs HP14.5 were divided in 3 groups: DMEM group， CMFb group and CMAFb group. MTT was used to draw the curves of cell growth. Immunofluorescence staining and real-time quantitative polymerase chain reaction (Real-time PCR) were performed for detecting the albumin (ALB)， alphafetoprotein (AFP) and cytokeratin18 (CK18) in protein and mRNA expression level. And the PAS staining was used to detect Glycogen synthesis. In the results， MTT showed faster proliferation of liver stem cells in CMAFb group compared with other groups; The Real-time PCR showed that the mRNA expression of ALB， AFP and CK18 in CMAFb group were much higher than in CMFb group and DMEM group after 5 days culture， while the number alpha-fetoprotein and mRNA expression level declined remarkably. Also， the similar results were observed in immunofluorescence. And the result of PAS staining suggested that the Glycogen synthesis in CMAFb group was much higher than the other two groups. Therefore， this research highly suggests that the fibroblasts activated by Hepa1-6 are able to increase the proliferation of EHSCs and differentiate the functional mature liver cells.

CSTR: 32200.14.cjcb.2014.07.0011