人肝细胞L02诱导MSCs分化为肝样细胞的观察  

Human bone marrow mesenchymal stem cells differentiating into hepatocyte-like cells after being induced by co-culturing with human hepatocyte line L02

作　　者：张立婷[1] 陈红[1] 冯利[2] 吴玉卓[1] 

机构地区：[1]兰州大学第一医院,甘肃兰州730000 [2]兰州大学

出　　处：《山东医药》2008年第24期4-6,共3页Shandong Medical Journal

基　　金：兰州大学医学基金资助项目(LZUYX200655)

摘　　要：目的探讨骨髓间充质干细胞(MSCs)在与肝细胞L02在体外共培养条件下诱导分化为肝细胞的可行性。方法密度梯度离心联合贴壁筛选法获取MSCs。将接种于盖玻片上的MSCs和人肝细胞L02共置于培养皿中。在2、4、6、8d时分别用免疫细胞化学检测AFP、细胞角蛋白(CK18),同时检测染色体核型。结果成功分离培养出MSCs,其表型为CD29阳性,CD34阴性。共培养的MSCs可在2～8d时,形态变为三角形、多角型或类圆形。第2天时,共培养细胞AFP表现为强阳性表达,以后减弱,第8天AFP的阳性表达很少。第2、4天检测出CK18均为阴性表达,第6、8天CK18为阳性表达。阳性对照组细胞CK18有较强的阳性表达,AFP弱阳性表达。阴性对照组均为阴性表达。共培养细胞染色体核型无融合现象。结论人肝细胞L02能够诱导MSCs分化为肝样细胞。Objectives To explore the possibility of Human bone marrow mesenchymal stem cells （MSCs） differentiating into hepatocyte-like cells by co-culturing with human hepatocyte line L02 in vitro. Methods MSCs were separated by density gradient centrifugation and were cultured through adhere culture. MSCs and L02 hepatocytes were spread on coverslips separately, and they were put in a culture dish. A separate culture of L02 hepatocyte served as a positive control and a separate culture of MSCs served as a negative control. The expression of AFP,CK18 in MSCs were detected by immunohistochemistry at days of 2,4,6,8. Karyotype was detected too. Results Growth and division of MSCs were good and the phenotypes of MSCs were CD29 positive and CD34 negative. The shape of MSCs changed into polygonal or round on days 2 - 8 in co-culturing groups. On the second day, the MSCs expressed a high level of AFP, then AFP expression tapered daily and there was hardly any expression of AFP on the day 8. The initial expression of CK18 appeared on day 6 and reached a higher level on day 8. The positive control cells had a high expression of CK18, while there was a weak expression of AFP. The negative control cells had no expressions for the two markers. MSCs were detected no chromosome fusion. Conclusion Human MSCs can be induced to differentiate into mature hepatocyte-like cells by co-culturing with L02 hepatocytes.

关 键 词：肝细胞 细胞分化 间充质干细胞 肝样细胞 

分 类 号：Q21[生物学—细胞生物学]