胆汁化血清对骨髓间充质干细胞向肝细胞样细胞定向分化的诱导

目的:寻找生物人工肝和肝细胞移植需要合适的肝细胞来源,研究不同诱导方法对骨髓间充质干细胞(MSC)在体外分化为肝细胞样细胞的影响。方法:本研究应用贴壁法对MSC进行分离和培养后,应用胆汁化血清及肝细胞生长因子不同诱导方式对之进行诱导分化,研究MSC转化为肝细胞样细胞的可能性,并进行转化的鉴定。结果:经贴壁分离和培养的MSC在5%胆汁化血清诱导作用下,21 d后可分化为形态似肝细胞样的细胞,经免疫组织化学显示,该细胞可表达细胞角蛋白18(CK18)和甲胎蛋白(AFP),且原位杂交显示,在培养的第35天,部分细胞还可合成清蛋白,与肝细胞生长因子(HGF)有着相似的诱导效果。结论:在合适的条件下,MSC可向肝细胞方向转化,且本方法操作简捷,胆汁化血清易于获得,具有潜在的应用价值。

Expansion and hepatocytoid cell induction of rat bone marrow mesenchymal stem cells in vitro

Objective To find suitable sources of hepatocyte for bioartificial liver and liver transplantation,and to investigate the possibility of bone mesenchymal stem cells(MSCs) directional differentiation into hepatocytoid cells.Methods MSCs were isolated and cultured by Friedensteion's method,then these cells were induced for the differentiation of hepatocytoid cells by exposure to the hepatocyte growth factor(HGF)or cholestatic serum,respectively.Results After 21 d of induction,the MSCs demonstrated cuboidal morphology,which was the characteristic of hepatocytes,and these cells were further confirmed to have cytokeratin 18(CK18) and AFP expression by immunohistochemistry staining.After 35 d,some of the induced cells demonstrated positive albumin mRNA expression.The induction efficacy of cholestatic serum was similar to that of HGF.Conclusions Rat MSCs can differentiate into hepatocytoid cells under appropriate condition.The culturation and induction procedures are easy to handle.This technique has potential value in the treatment of liver failure.