The Differentation Of Homogeneity And Allogeneic Rats Bone Marrow Mesenchymal Stem Cells Into Liver-like Cells In Vitro

Objective: We observe the general biological features and immunogenicity of Mesenchymal stem cells (Mesenchymal stem cells, MSCs) and explore whether the homogeneity and allogeneic rats bone marrow MSCs can be differentiate into liver-like cells in vitro induced by HGF and FGF-4 .The purpose of this study is to provide a new thoughts for the homogeneity and allogeneic adult bone marrow MSCs therapy for end-stage liver disease.Method:1.In Aseptic conditions, the homogeneity and allogeneic rats bone marrow MSCs were collected sterilely from a healthy SD rats weighted about 120-150g, aged 6-8 weeks , then MSCs were cultivated, purified and proliferated in DMEM-LG Medium containing 10%FBS by adherence wall cell culture technique.2.MSCs from the third generation by flow cytometry to testCD34,CD45,CD29,CD106 and induce MSCs into fat cells.3.Take two third-generation rat MSCs mixed culture by adding a certain concentration of HGF and FGF-4 on MSCs induced by training, did not join the group as a growth factor in the control group to observe the morphological changes induced cells, immunohistochemical detection of training-induced cells Cytokeratin -18 (cytokeratin-18 CK-18), cytokeratin -19 (cytokeratin-19 CK-19) and vimentin Vimentin; ELISA detection methods such as alpha-fetoprotein (alpha-fetoprotein AFP), albumin (albumin ALB), alkaline phosphatase (alkaline phosphatase ALP).Results:1.freshly isolated bone marrow MSCs had uniform size and form,and transparency was perfect.MSCs anhered to the cultured flask wall within 24hours and adherence cell number increased in clonal manner as time went on. Spindle or triangular shaped cells appeared at day 3 and reached logarithmic stage by day 10.At passage 3,MSCs fused into single layer and arranged in distinct directions so that they looked like swirl,shoal,reticulation.2.The third generation of rat MSCs by flow cytometry CD34,CD45 are negative, CD29, CD106 are positive, and they were induced to differentiate into fat cells.3.After being culture with HGF and FGF-4,the MSCs showed multilateral and ovlivary transformation. The differentiatiated cells expressed albumin,AFP,CK18, CK19and vimentin.Conclusions:1.HGF and FGF-4 can induce the allogeneic rat MSCs to differentiate into hepatocyte-like cells.2.DMEM low glucose medium containing 10% fetal bovine serum was suitable for culture and proliferation of rat bone marrow MSCs in vitro.3.Bone marrow-derived mesenchymal stem cells are low immunogenicity.4.Bone marrow-derived MSCs could be a liver tissue engineering major source of seed cells.