胰岛与Sertoli细胞体外共同培养

Coculture of rat islets and Sertoli cells in vitro

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 investigate a new method of a long term culture for rat islets in vitro. Methods Rat islets marked by green fluorescent protein (GFP) were cocultured with Sertoli cell for 20 weeks. Histological studies were performed on islet group and coculture group in 1w, 3w, 10w, 15w, 20w of culture by light, fluorescent and electron microscopy. Insulin released was measured by radioimmunoassay. Results In islet group, islet viability and the number of insulin positive cells were significantly decreased after 3w of culture, cumulative quantities of insulin for 24 hours and the stimulation index also fell rapidly under this condition, meanwhile the ultrastructure of islets was destroyed. However, under coculture condition, culture time of islets was prolonged in vitro, islet viability and the number of insulin positive cells were significantly increased, cumulative quantities of insulin for 24 hours and the stimulation index maintained at high level, and the ultrastructure of islets remained normal even after 20w of culture. Conclusion Coculture of rat islets with Sertoli cells may promote islet growth and prolong culture time, and it is a new method of a long term culture of islets in vitro.