目的 研究体外共培养模型中肌源性干细胞(MDSCs)改善大鼠胰岛活性的作用.方法 提取新生大鼠MDSCs原代细胞,差速贴壁培养后行结蛋白免疫细胞化学鉴定,取第4代MDSCs接种在transwell底板.提取大鼠胰岛后,双硫腙染色鉴定,胰岛在高糖环境中处理24 h后放入tran-swell板上层与MDSCs共培养.在培养的第1、3、6天分别用吖啶橙/溴化乙锭(AO/EB)双荧光染色胰岛,RT-PCR检测胰岛的Bcl-2/Bax基因表达来评价胰岛凋亡,其结果与单独胰岛培养组比较.结果 与单独胰岛培养相比,共培养组第3、6天AO/EB染色见胰岛细胞明显增多,Bcl-2明显升高,而Bax明显降低(P<0.05、P<0.01).结论 大鼠胰岛与MDSCs共培养延长大鼠胰岛存活时间,改善胰岛活性.%Objective To study the effect of co-culture in vitro with muscle-derived stem cells (MDSCs) on the function of the pancreatic islets in rat model. Methods MDSCs were extracted by mixed enzymatic digestion and purified by differential adherent culture, which were identified with Desmin immunocytochemical staining. MDSCs of the fourth generation were inoculated in the bottom room of transwell. Rat pancreatic islets were extracted and then identified by DTZ dyeing. After the islets had been brought up in high sugar situation for 24 hours,they were added in the upper room of transwell. The islets were dyed by AO/EB on the 1st, 3rd, 6th day, respectively. The expression levels of Bcl-2 and Bax mRNA in these cells were measured by semi-quantitive RT-PCR to evaluate the apoptosis of islet cells. The results were compared to those from the islets cultured without MDSCs. Results Compared with cultured without MDSCs,the number of AO dyeing islet cells in co-cultured group was increased obviously on the 3rd and 6th day. The expression of Bcl-2 increased and Bax mRNA decreased on the 3rd and 6th day in co-cultured group (P＜0.05 and P＜0. 01). Conclusion Rat pancreatic islets live for more time in the co-culture group than those in the separately cultured group and the viability of islets is improved in the co-culture group.
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