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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 investigate the effect of hepatocyte growth factor (HGF) combined with fibroblast growth factor-4 (FGF-4) on the induction of human bone marrow mesenchymal stem cells The ability of hepatocytes to differentiate. Methods HMSCs were induced to differentiate into hepatocyte-like cells by HGF, FGF-4 and HGF + FGF-4 respectively. AFP and cytokeratin 18 (CK18) were detected by immunocytochemical staining at different stages of differentiation. AFP and albumin (ALB) were detected by immunofluorescence staining ; RT-PCR detection of AFP and ALB mRNA expression. Results HMSCs were induced to transform into hepatocyte-like cells. Immunocytochemical staining of each induction group can detect AFP and CK18 expression. The results of image analysis showed that the positive rate of AFP and CK18 induced by HGF + FGF-4 was the highest, followed by HGF and FGF-4 was weaker. Immunofluorescence staining showed that AFP and ALB were positive in each induction group The expression of AFP and ALB mRNA was detected by RT-PCR, but not in the negative control group. Conclusions HGF, FGF-4 and HGF + FGF-4 can induce HMSCs to differentiate into hepatocyte-like cells. Differentiated hepatocyte-like cells can secrete hepatocyte-specific products AFP, ALB and CK18. HGF + FGF-4-induced HMSCs differentiated into liver-like cells the highest positive rate.