摘要
目的探讨诱导人脐血间充质干细胞(UBCMSCs)向类肝细胞定向分化的可能性。方法采用密度梯度离心法与直接贴壁法相结合,从人脐血中分离UBCMSCs;通过流式细胞仪检测细胞表面抗原CD34、CD44,确定UCBMSCs的比例。在UCBMSCs培养体系中,加入人肝细胞株L-02细胞培养上清液或诱导因子(联用HGF和FGF-4)共培养,检测诱导后第7天、14天细胞内细胞角蛋白18(CK18)、白蛋白及糖原的表达。结果从脐血中分离获得均一贴壁的UBCMSCs;在UBCMSCs培养体系中,无论是添加L-02细胞培养上清液,还是采用诱导因子HGF和FGF-4,均可诱导UCBMSCs细胞内表达中晚期成熟肝细胞表面标志物CK8&18、白蛋白及糖原。结论密度梯度离心法和直接贴壁法相结合能够有效分离UCBMSCs;L-02人肝细胞培养上清液或诱导因子HGF、FGF-4均具有诱导UCBMSCs向类肝细胞定向分化的作用。
Objective To evaluate the possibility of human umbilical cord blood mesenchymal stem ceils （UCBMSCs） in inducing to differentiate into hepatocyte-like cells in vitro. Methods The UCBMSCs were isolated by density gradient centrifugation and directly adherence growth, then treated with supernatant fluid of human hepatocyte line L-02 cultivation or HGF/FGF-4.7 days and 14 days later,those ceils were collected for detecting CK18, albumin and glycogen by immunohistochemistry;Meanwhile,CD44 and CD34 were detected by flow cytometry. Resalts The adherent UCBMSCs were obtained and began to express CK8＆18,human albumin and glycogen after co-cultivation with supernatant fluid of human hepatocyte line L-02 or HGF/FGF-4. Conclusions UCBMSCs can be obtained by density gradient centrifugation and directly adherence growth. They can also differentiate into hepatocyte-like ceils which express CK8＆18, albumin and glycogen in some conditions. It provides a powerful model for further study of cellular senescence and for future application to cell-based therapy by using umbilical cord blood ceils.
出处
《中国基层医药》
CAS
2006年第12期1976-1979,共4页
Chinese Journal of Primary Medicine and Pharmacy
关键词
脐血
间充质干细胞
肝细胞
Umbilical cord blood
Mesenchymal stem cells
Hepatocyte
