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Objective: To establish a human bone marrow mesenchymal stem cells ( mesenchymal stem cells , MSCs ) cultured in vitro method to explore the hepatocyte growth factor ( HGF ) , epidermal growth factor ( EGF ) - induced differentiation of MSCs into hepatocyte- directed feasibility . Method : using Ficoll lymphocyte separation liquid isolated from the bone marrow MSCs were seeded in the L - DMEM culture medium containing 10% fetal bovine serum were cultured and passaged the the pending basic cell fusion mixture was digested with trypsin and EDTA . Cell morphology was observed under inverted phase contrast microscope , trypan blue staining to count the cells and growth curve , flow cytometry cell cycle . Take the first three generations of MSCs with HGF , EGF , HGF EGF and growth factor - free serum-free liver cell culture medium induced culture . Morphological changes were observed under inverted phase contrast microscope , 1,7,14,21,28 induction remove cells seeded immunocytochemistry were used to detect alpha-fetoprotein (AFP), albumin ( ALB ) expression of cytokeratin 18 ( CK18 ) , PAS law the glycogen staining test, and the culture supernatant automatic biochemical analyzer ALB and BUN content . Results: 1 by density gradient centrifugation combined with adherent culture method can be successfully isolated from human bone marrow MSCs . Just inoculated cell morphology was circular, triangular or coryneform after 48 h , cells begin to adherent ,4 - 5d visible colony formation , the formation of multiple cell clones 7d , 10d fusiform cells , up to about 2W basic integration ; passage cells were uniform spindle-shaped , basically no suspension cells . The growth curve of growth slow demurrage , the typical characteristics of the logarithmic growth phase and growth of the stationary phase . Flow cytometry showed that about 86.4 % of the cells in the G0, G1 phase . Inoculated containing HGF , MSCs of EGF and HGF the EGF No serum liver cell culture medium in the induction of the fifth day can be observed in the part of the spindle-shaped cells became round , a slight decrease in the number of cells in the medium was changed at the same time inducing hole ; 10 days basic
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