肝细胞生长因子诱导骨髓间充质干细胞表达甲胎蛋白和白蛋白

Expression of alpha fetoprotein and albumin in bone marrow mesenchymal stem cells induced by hepatocyte growth factor

背景:对于诱导骨髓间充质干细胞成肝细胞样细胞的研究,在大鼠及小鼠及人类中已有相关报道。甲胎蛋白和白蛋白均为肝细胞系较为特异的标志。目的:进一步验证肝细胞生长因子对骨髓间充质干细胞的诱导分化作用。方法:采用密度梯度离心和贴壁培养法相结合分离培养人骨髓间充质干细胞,以含20μg/L肝细胞生长因子和10μg/L碱性成纤维细胞生长因子的低糖DMEM培养基诱导培养,对照组取同代细胞,常规培养液培养。诱导培养7,14,21和28d采用免疫组织化学方法检测甲胎蛋白和白蛋白的表达及RT-PCR技术检测白蛋白mRNA表达。结果与结论:诱导组骨髓间充质干细胞呈类上皮样细胞。诱导组在培养7d时,甲胎蛋白表达阳性率为81.5%,随着培养时间延长,甲胎蛋白阳性表达率逐渐减弱,而白蛋白阳性表达率及白蛋白mRNA表达逐渐增强,至培养28d,白蛋白阳性表达率达91.2%,对照组细胞均无白蛋白、白蛋白mRNA及甲胎蛋白表达。结果证实,肝细胞生长因子可诱导骨髓间充质干细胞表达甲胎蛋白和白蛋白。

BACKGROUND：Reports have addressed that induction of bone marrow mesenchymal stem cells（BMSCs） into hepatocyte-like cells in rats,mice and human.Alpha fetoprotein and albumin are specific markers of hepatocyte line.OBJECTIVE：To verify induction effects of hepatocyte growth factor on differentiation of BMSCs.METHODS：Human BMSCs were isolated and purified by density gradient centrifugation and adherent culture.BMSCs were cultured in low glucose-DMEM containing 20 μg/L hepatocyte growth factor and 10 μg/L basic fibroblast growth factor in vitro.The same passage of cells in the control group was selected and cultured in routine medium.The expression of alpha-fetoprotein and albumin was detected by immunohistochemistry at 7,14,21 and 28 days.The expression of albumin mRNA was detected by RT-PCR.RESULTS AND CONCLUSION：BMSCs became epithelia-shaped cells in the induction group.Alpha fetoprotein-positive rate was 81.5% at day 7 in the induction group.With prolonged time,alpha fetoprotein-positive expression became weak,whereas albumin-positive rate and albumin mRNA expression became strong.Albumin-positive expression rate was 91.2% by 28 days.In the control group,no albumin,albumin mRNA and alpha fetoprotein expression was determined.Results have verified that hepatocyte growth factor can induce BMSCs expressing alpha fetoprotein and albumin.