体外定向诱导脐血间充质干细胞向类肝细胞分化的研究

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。

Differentiation of mesenchymal stem cells in umbilical blood into hepatocyte-like cells in vitro

Objective To evaluate the possibility of human umbilical cord blood mesenchymal stem cells(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 cells were collected for detecting CK18,albumin and glycogen by immunohistochemistry;Meanwhile,CD44 and CD34 were detected by flow cytometry.Results 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 cells 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 cells.