肝细胞生长因子诱导人骨髓来源的多能成体祖细胞向肝样细胞分化的可行性

目的探索人骨髓来源多能成体祖细胞(mulfipotent adult progenitor cells from human bone marrow，hMAPCs)在一定诱导条件下分化为肝细胞的可行性，为肝组织工程提供新的种子细胞来源。方法分组：A组1(hepatocyte growth factor，HGF)20ng／mL诱导；B组(阳性对照组)：L-02人肝细胞株；C组(阴性对照组)：未加任何诱导因素的MAPCs。免疫细胞化学鉴定不同诱导分化阶段细胞的白蛋白(albumin，ALB)、甲胎蛋白(alpha fetoprotein，AVP)、细胞角蛋白18(cytokeratin 18，CK-18)、细胞角蛋白19(cytokeratin 19，CK-19)等肝细胞特征的表型变化并计数阳性细胞比率；Western blot检测诱导分化第21、35天后细胞的ALB的表达。结果免疫细胞化学结果：ALB、CK-18在不同时间段基本为阳性着色；CK-19在各组均为阴性着色，AFP作为早期不成熟肝细胞的一个标志，仅在诱导第7天为阳性着色；Western blot检测结果：在诱导分化第21、35天，ALB均有阳性表达。结论hMAPCs在一定诱导条件下具有向肝样细胞分化的潜能。

Investigation possibility of multipotent adult progenitor cells from human bone marrow differentiating into hepatocytes in vitro

[Objective]To investigate the possibility of the Multipotent adult progenitor cells from human bone marrow(hMAPCs) which are induced into hepatocytes with Hepatocyte growth factor (HGF) in vitro, and find a new cell source for liver tissue engineering. Divided groups: A: induced with HGF (20 ng/mL); B: control group: L-02 human hepatocytes(cell lines), C: contrast: the undifferentiated hMAPCs. The expressions of Albumin(ALB), Alpha Fetoprotein (AFP), Cytokeratin 18(CK-18), Cytokeratin19(CK-19) were detected with immunocytochemistry to identify the characters of the differentiated cells in different time and determed the ratio of positive cells at 7, 14, 21 days. ALB expressions of the differentiated cells in different time were detected by Western blot at 21, 35 days. The result of immunocytochemistry:A groups cells expressed the late markers (ALB, CK-18) of hepatocyte in all the different times. The cells of all groups did not express CK-19. As a early marker(AFP) of hepatocyte, A groups expressed AFP in early differentiation time, and did not expressed AFP at late differentiation time. The result of Western blot assay: the differentiated cells expressed the late markers(ALB) of hepatocyte on day 21, 35. [Conclusions] hMAPCs under special inducing conditions can differentiate into hepatocyte-like cells, which provide us a potential cell source for liver tissue engineering.