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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 investigate the effect of myogenic stem cells (MDSCs) on rat pancreatic islet activity in vitro co-culture model. Methods Primary cultured neonatal rat (MDSCs) primary cells were isolated and identified by immunocytochemistry after differential adherent culture. Generation 4 MDSCs were seeded on transwell plates. The rat islets were extracted and identified by dithizone staining. The islets were placed in the supernatant of tran-swell plate and cultured with MDSCs for 24 h in a high glucose environment. On the 1st, 3rd and 6th day of culture, the pancreatic islets were double-stained with acridine orange / ethidium bromide (EB) and the expression of Bcl-2 / Bax was detected by RT- Results compared with islet culture alone. Results Compared with the islet culture alone, AO / EB staining on the 3rd and 6th day in the co-culture group showed that the number of islet cells, Bcl-2 and Bax were significantly decreased (P <0.05, P <0.01). Conclusion Co-culture of rat islets with MDSCs prolongs islet survival and improves pancreatic activity.