【摘要】 目的探讨人参皂甙(CS)与肝细胞生长因子(HGF)体外诱导骨髓间质干细胞(MSC)分化为肝细胞的协同效应及分子机制。方法采用 Ficoll-Paque 液离心分离鼠 MSC,体外扩增,在 LDMEM 培养液中加入 HGF 和 GS 进行诱导分化。通过细胞形态学观察、免疫组化检测白蛋白、CK8/18的表达以及 RT-PCR 检测白蛋白、CK18等肝细胞特征性蛋白的 mRNA 转录水平、糖原染色等来分析诱导分化的效果。结果鼠MSC 体外扩增3代,细胞数可达(1～2)×10~7左右。加入 HGF 和 GS 诱导后,MSC 形态逐渐分化为肝细胞样,免疫组化染色见胞浆内出现棕黄色颗粒,表达肝细胞特有的白蛋白以及 CK8/18等蛋白。RT-PCR 显示有白蛋白、CK18等的 mRNA 转录。糖原染色亦呈阳性反应。结论人参皂甙与肝细胞生长因子在体外可以诱导骨髓间质干细胞分化为肝细胞样细胞。更多还原

【Abstract】 Objective To observe the synergistic effect and molecular mechanism of Ginsenosides(GS)and hepatic growth factor in inducing the differentiation of mesenchymal stem cells(MSC)into hepatocytes in vitro.Methods Ficoll-Paque liquid was adopted to centrifuge mouse MSC and expand in vitro.Hepatic growth factor(HGF)and GS were added into L- DMEM cultural medium to induce cell differentiation.Albumin,CK8/18 expression and staining for glycogen were deter- mined by cytomorphological observation and immunochemical method.mRNA transcription level of albumin and such hep- atocyte as CK18 were analyzed by RT-PCR method.Results MSC of rats were expanded into 3 generations,with the cell number reaching(1-2)×10~7.The morphology of MSC was gradually differentiated into the hepatocellular phenotype when induced by adding HGF and GS.Brown granules were seen in cytoplasm when immuochemical staining,expressing the peculiar characters of albumin and CK8/18 protein of hepatocytes,mRNA tmscription of albumin and CK18 was shown by RT-PCR.Positive reaction was also shown in glycogen staining.Conclusion GS and HGF could induce the differentiation of bone marrow mesenchymal stem cells into hepatocellular phenotype.更多还原