人骨髓间充质干细胞体外分化为肝细胞样细胞

The differentiation of human bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro

目的 探讨人骨髓间充质干细胞(MSCs)的体外培养及特异性诱导为肝细胞样细胞的能力。方法 骨髓标本来源于健康志愿者的胸骨,年龄2～35岁,采用淋巴细胞分离液(密度1.077)分离人MSCs,并分别采用HGF、FGF4、HGF+FGF4以及无生长因子四种处理因素体外诱导第三代人MSC向肝细胞样细胞分化。通过流式细胞术分析鉴定.MSCs的纯度,于诱导培养的0、7、14、21、28天时留取细胞检测CK18、AFP和白蛋白的表达情况,同时进行糖原染色验证细胞功能。结果 用淋巴细胞分离液分离出的人MSCs纯度可达90％,采用HGF、FGF4及HGF+FGF4三种处理因素均可在体外诱导人MSCs特异分化为具有肝细胞样细胞表型和功能的细胞。结论 人MSCs能在体外扩增并定向诱导为肝细胞样细胞。

Objective To investigate whether human bone marrow mesenchymal stem cells can differentiate into hepa-tocyte-like cells in vitro. Methods Bone marrow cells were collected from the sternum of healthy volunteers aged from 2 to 35,hMSCs were separated with Lymophocyte Separating Liquid (density 1.077). HGF , FGF4 , HGF + FGF4 was added respectively to induce the differentiation progress of hMSCs after passagez.The purity of hMSCs was analyzed by flow cytometry, the expressions of CK18,AFP and albumin were detected on day (0.7,14,21,28 after induction, and function of hepatocyte-like was tested by PAS (Periodic acid-schiff)after induction.Results The purity of hMSCs was 90% after separated by Lymophocyte Separating Liquid as assessed by flow cytometry, hMSCs could differentiate into cells with hepatic phenotype and function in the presence of HGF and (or) FGF4 in vitro.Conclusion hMSCs can expand and differentiate into hepatocyte-like cells in vit-