大鼠骨髓间充质干细胞体外诱导为肝细胞样细胞

Induced differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro

背景:目前体外诱导骨髓间充质干细胞分化为肝细胞样细胞的研究主要集中在不同诱导因子的诱导作用,而微环境的诱导作用研究较少。目的:观察肝细胞生长因子、胎肝细胞对骨髓间充质干细胞体外诱导分化为肝细胞样细胞的影响。方法:采用密度梯度离心法分离大鼠骨髓间充质干细胞,反复贴壁法纯化扩增骨髓间充质干细胞;采用Ⅰ型胶原酶消化法分离孕3周大鼠胚胎肝脏细胞,差速贴壁法纯化肝脏细胞;阴性对照组骨髓间充质干细胞只加入含体积分数为10%胎牛血清的L-DMEM培养基。诱导组在阴性对照组基础上加入一定浓度肝细胞生长因子或者与胎肝细胞共培养进行诱导分化。结果与结论:诱导组白蛋白、甲胎蛋白水平均高于非诱导组(P<0.01),诱导组糖原染色阳性,免疫细胞化学染色CK-18阳性,而非诱导组糖原染色、CK-18均阴性。结果显示肝细胞生长因子和胎肝细胞均可在体外诱导大鼠骨髓间充质干细胞分化为具有肝细胞样细胞表型和功能的类肝细胞。

BACKGROUND：The study of the induction of differentiation of rat bone marrow mesenchymal stem cells（BMSCs） into hepatocyte-like cells majorly concentrates on the induction of different inducing factors,and pays little attention on the induction of micro environment.OBJECTIVE：To investigate the differentiation of rat BMSCs into hepatocyte-like cells.METHODS：The rats BMSCs were isolated by density gradient centrifugation and purified by adherence cultivation,morphology feature of BMSCs were observed by microscope.The fetal liver cells were isolated by adherence cultivation after digesting the rats embryo liver of 3 weeks with collagenase.The negative control group was cultured in L-DMEM containing 10% fetal bovine serum.The purified BMSCs in the induced group were cultured in L-DMEM containing 10% fetal bovine serum with hepatocyte growth factor（HGF） and fetal liver cells.RESULTS AND CONCLUSION：Compared with those in non-induced BMSCs,the levels of alpha-fetoprotein and albumin in the induced-BMSCs were higher（P〈0.01）.Both glycogen and CK-18 were positive in the induced BMSCs.BMSCs can differentiate into hepatocyte-like cells with hepatic phenotype and function in the presence of HGF and fetal liver cells,which may be used as a kind of cell resources to treat severe hepatic disease.