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目的 研究胰岛体外长期培养的新方法。方法 采用绿色荧光蛋白 (GFP)标记成年SD大鼠胰岛 ,与Sertoli细胞共同培养 2 0周。应用形态学方法和透射电镜观察胰岛细胞单独培养和共同培养的生长特性 ;放射免疫法测定胰岛素分泌量。结果 胰岛单独培养 3周 ,细胞存活率显著下降 ,胰岛素阳性细胞数目减少 ,胰岛素 2 4h累积分泌量和对葡萄糖刺激的反应性下降 ,大部分胰岛 β细胞超微结构破坏 ,分泌颗粒减少 ;与Sertoli细胞共同培养的胰岛存活时间显著延长 ,细胞存活率和胰岛素阳性细胞数目较单独培养显著增加 ,胰岛素分泌量始终处于高水平状态 ,培养 2 0周胰岛 β细胞超微结构基本正常。结论 大鼠胰岛与Sertoli细胞共同培养可以促进胰岛生长 ,显著延长存活时间 ,是一种新的体外长期培养胰岛的方法。
Objective To study a new method of islet in vitro long-term culture. Methods The adult SD rat islets were labeled with green fluorescent protein (GFP) and cultured with Sertoli cells for 20 weeks. Morphological methods and transmission electron microscopy were used to observe the growth characteristics of islet cells cultured alone and co-cultured. The amount of insulin secretion was measured by radioimmunoassay. Results After islet cultured for 3 weeks alone, the cell viability decreased significantly, the number of insulin positive cells decreased, the cumulative secretion of insulin at 24 h and the reactivity to glucose stimulation decreased, the ultrastructural destruction of most islet β cells and the secretion of granules decreased; The co-cultured islets survival time was significantly longer, the cell survival rate and the number of insulin-positive cells were significantly increased compared with the culture alone, the insulin secretion was always at a high level, and the ultrastructure of islet β cells was basically normal after 20 weeks of culture. Conclusion Co-culture of rat pancreatic islets and Sertoli cells can promote the growth of islets and prolong the survival time significantly. It is a new method of long-term culture of islets in vitro.