摘要
目的探讨肝细胞生长因子(hepatocyte growth factor,HGF)联合成纤维细胞生长因子-4 (fibroblast growth factor-4,FGF-4)诱导人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,HMSCs)向肝细胞方向分化的能力。方法分别用HGF、FGF-4、HGF+FGF-4诱导HMSCs分化为肝样细胞。在不同分化阶段用免疫细胞化学染色法检测甲胎蛋白(alpha fetoprotein,AFP)和细胞角蛋白18(cytokeratin 18,CK18);免疫荧光细胞化学染色法检测AFP、自蛋白(albumin,ALB)的表达; RT-PCR检测AFP和ALB mRNA表达的情况。结果HMSCs经诱导向肝样细胞转化。免疫细胞化学染色各诱导组均可检测出AFP和CK18表达。图像分析表明HGF+FGF-4联合诱导分化的AFP、CK18阳性率最高,HGF次之,FGF-4则较弱;免疫荧光细胞化学染色发现各诱导组AFP、ALB均为阳性表达;各诱导组RT-PCR均可检测出AFP和ALB mRNA的表达,而阴性对照组则没有表达。结论HGF、FGF-4、HGF+FGF-4均能诱导HMSCs分化为肝样细胞,分化的肝样细胞能分泌肝细胞特异性产物AFP、ALB、CK18等。以HGF+FGF-4诱导HMSCs分化为肝样细胞的阳性率最高。
Objective To induce human bone marrow mesenchymal stem cells （HMSCs） differentiation into hepatocyte-like cells by using hepatocyte growth factor （HGF） plus fibroblast growth factor-4 （FGF-4） in vitro. Methods HMSCs were induced to differentiate into hepatocyte-like cells by HGF, FGF-4 and HGF ＋ FGF-4 in vitro. The expressions of hepatic markers, AFP and CK-18, were detected by immunocytochemical staining at different times after induction. The expressions of hepatic markers, AFP and ALB, were detected by immunofluorescence in each group at different times after induction. The expressions of AFP, ALB mRNA were detected by RT-PCR assay after induction. Results HMSCs transformed into hepatocyte-like cells gradually after induction. The expressions of AFP and CK18 were positive after induction in group B, C, D, E as detected by immunocytochemical staining. The positive rates of AFP and CK18 in group D in which HGF and FGF-4 were added as calculated with image analysis were significantly higher than that in group B, C and undifferentiated group. The expressions of AFP, ALB were both positive after induction in each group except group A. Similarly, the expressions of AFP and ALB mRNA were both positive as detected by RT-PCR after induction in each group except group A. Conclusion HMSCs were induced to differentiate into hepatocyte-like cells by HGF, FGF-4 separately or HGF ＋ FGF-4 in combination, which express some hepatic markers such as AFP, ALB and CK18. The effect of HGF ＋ FGF-4 is among the strongest.
出处
《中华普通外科杂志》
CSCD
北大核心
2006年第11期780-782,共3页
Chinese Journal of General Surgery
基金
广东省科技计划项目(2002A3020206)
关键词
间质干细胞
细胞分化
肝样细胞
Mesenchymal stem cells
Cell differentiation
Hepatocyte-like cells
