瘀胆血清体外诱导骨髓间充质干细胞向肝细胞分化的实验研究论文.docVIP

　　瘀胆血清体外诱导骨髓间充质干细胞向肝细胞分化的实验研究论文 钟晓琳，万居易，王忠琼，李昌平，胡莲 【摘要】 目的 探讨骨髓间充质干细胞(MSCs)体外诱导其分化为肝细胞的可行性。方法 分离、纯化并鉴定大鼠骨髓MSCs，制备正常及瘀胆大鼠血清，取第3代MSCs分组培养：A组：DMEM+10%FBS;B组：肝细胞生长培养基(HGM)+5%FBS; C组：HGM+5%正常大鼠血清;D组：HGM +5%瘀胆大鼠血清;E组：HGM +5%瘀胆大鼠血清+25μg/L HGF。观察各组细胞形态变化，MTT法测定细胞生长曲线.freelRNA阳性表达，第14，21天出现CK18 mRNA阳性表达，D、E组上清液中白蛋白浓度随诱导时间延长逐渐升高，且E组高于D组(P 0.05)。A、B、C组未见AFP、CK18表达及白蛋白浓度变化。结论 正常大鼠血清能明显促进MSCs的增殖;瘀胆血清对MSCs有一定促增殖作用，且能有效诱导其向肝细胞分化，联合使用HGF能提高分化率。 【关键词】 瘀胆血清;肝细胞生长因子;间充质干细胞;肝细胞 Research on the Differentiation of Mesenchymal Stem Cells Induced by Chelestatic Sera in vitro into Hepatocyte Zhong Xiaolin, edical Colleg, Luzhou City, Sichuan Province 646000 P.R.China Abstract Objective To discuss the possibility of the differentiation of mesenchymal stem cells (MSCs) induced by chelestatic sera in vitro into hepatocyte. Methods Rat bone marroal and cholestatic sera of rats edium(HGM)+5%FBS), Group C (HGM+5% normal rat sera), Group D (HGM+5% cholestatic rat sera) and Group E (HGM+5% cholestatic rat sera+25 μg/L HGF); The morphologic changes of the induced cells easured erase chain reaction (RT-PCR) and the albumin content in supernatant ined by bromocresol green method.Result MSCs polygonal and binuclear cells orphologic changes of the cells onstrated that the cells in Group C proliferated the fastest and those in Group D and Group E proliferated faster than those in Group B (P 0.05); the positive expression of AFP m RNA RNA in concentration in these 2 groups increased along e length of induction, and the concentration in Group E in concentration changes al rat sera can obviously promote the proliferation of MSCs; cholestatic sera is of certain effect in MSCs proliferation and can effectively induce the differentiation of MSCs into hepatocytes, the bined application esenchymal stem cells (MSCs) hepatocyte 近年来的研究表明，骨髓间充质干细胞(Mesenchymal stem cell，MSCs)在一定条件下能分化为肝细胞1，这为临床上解决肝细胞移植、生物人工肝等的肝源短缺问题提供了新的思路2，但MSCs的分化方向受周围环境多种因素影响和决定，寻求适合MSCs增殖及向肝细胞定向分化的条件是目前尚需解决的重要问题。本研究采用含正常大鼠血清、瘀胆血清及肝细胞生长因子(he