论文部分内容阅读
目的体外诱导分化骨髓间充质干细胞(mesenchymal stem cells,MSCs)为胰岛样细胞,鉴定诱导细胞并检测其功能。方法取周龄大鼠骨髓细胞,进行MSC培养纯化扩增,用bFGF、EGF、条件培养液、高糖培养液、B27、地塞米松、胰岛素等培养体系诱导MSC向胰岛细胞转化。双硫腙染色鉴定胰岛样细胞;免疫细胞化学鉴定胰岛素抗原表达;Western blot检测胰岛样细胞胰岛素的表达。结果体外诱导的MSCs由长梭形变成多角形并逐渐聚集成团。双硫腙染色细胞团成亮红色,初步表明为胰岛样细胞。诱导的细胞具有胰岛素抗原表达;Western blot检测诱导分化为胰岛样细胞的胰岛素表达阳性。结论骨髓间充质干细胞能够分化成胰岛样细胞,并具有分泌胰岛素的功能。
Objective To induce differentiation of mesenchymal stem cells (MSCs) into islet-like cells in vitro and to identify induced cells and test their function. Methods Bone marrow cells from week-old rats were harvested and purified by MSC culture. MSCs were transformed into pancreatic islet cells by bFGF, EGF, conditioned medium, high glucose medium, B27, dexamethasone and insulin. Dithizone staining was used to identify islet-like cells. Immunocytochemistry was used to identify the expression of insulin antigens. Western blot was used to detect the expression of insulin in islet-like cells. Results In vitro induced MSCs transformed from long fusiform to polygonal and gradually clustered into clusters. Dithizone staining cell mass into bright red, initially shown as islet-like cells. The induced cells had insulin antigen expression; Western blot showed that insulin induced positive differentiation into islet-like cells. Conclusion Bone marrow-derived mesenchymal stem cells can differentiate into islet-like cells and have the function of secreting insulin.