摘要目的 探讨间充质干细胞与胰岛联合培养在延长胰岛体外存活时间、保护胰岛功能方面的作用.方法 以4周龄Wistar大鼠作为供体,分离纯化骨髓间充质干细胞并培养鉴定;Histopaque-1077一步法分离纯化Wistar大鼠胰岛;将胰岛培养分为单纯胰岛基础培养、单纯胰岛高糖培养、胰岛与间充质干细胞联合基础培养、胰岛与间充质于细胞联合高精培养4组,每组又分3个时间段(3、7、14 d)观察胰岛形态变化及胰岛存活率,酶联免疫吸附法检测胰岛素分泌量及刺激指数.结果 骨髓间充质干细胞传代培养3代后,流式细胞术检测CD45及CD90,二者荧光强度差异有统计学意义(P＜0.05);联合培养组3、7、14 d的胰岛存活率均高于单纯培养组(P＜0.01);高糖刺激下联合培养组7 d的胰岛素分泌量及刺激指数均高于单纯培养组(P＜0.01).结论 间充质干细胞与胰岛联合培养可以明显延长胰岛体外存活时间并保持其活性,对胰岛具有良好的保护作用.

abstractsObjective To observe the effect of mesenchymal stem cells (MSCs) on enhancing rat islets viability and function in vitro by a pretransplant co-culture. Methods 4-week-old Wistar rats were used as donors, bone mesenchymal stem cells were isolated and subcultured. Islets of Wistar rats were isolated and purified by one-step single-layer Histopaque-1077. Then islets were divided into four groups randomly, 2 groups co-cultured with mesenchymal stem cells (MSCs) (one using low-glucose medium; the other using high-glucose medium ); 2 groups were cultured alone (low-glucose medium; high-glucose medium), each group was further stratified into 3 subgroups(3, 7, 14 d); the survival and functionality of these islets were observed and evaluated. The amount of glucose stimulated secreted insulin were measured wth a rat/mouse insulin enzyme-linked immunosorbent assay (ELISA) kit and stimulation index was also calculated. Results Compared with those not co-cultured, islets co-cultured with MSCs demonstrated significantly higher survival rates and viability both in 3th, 7th and 14th day ( P ＜ 0. 01 ); furthermore, cocultured islets revealed higher levels of glucose stimulated insulin secretion and secretion indexes in 7th day (P＜0.01). Conclusion Rat islet cells co-cultured with MSCs have longer in vitro survival and better functions.