大鼠胚胎成纤维细胞与胰岛共培养的实验研究

目的 探讨大鼠胚胎成纤维细胞(EFCs)与胰岛共培养对胰岛活性与功能的影响.方法 制备SD大鼠胚胎饲养层;分离SD大鼠胰岛.实验分为单纯胰岛培养组、EFCs与胰岛共培养组和EFCs单纯培养组.胰岛活性以AO/PI、DTZ染色、线粒体活性检测,以胰岛素释放试验评价其功能.结果 共培养组中,AO/PI双染显示胰岛活性高于95%,MTT显示共培养3、5、7、14d胰岛细胞吸光度明显高于单纯培养组(P＜0.05或P＜0.01),刺激指数SI也明显高于对照组.结论 新鲜分离的胰岛与EFCs饲养层共培养可明显改善胰岛功能.

IMPROVED ISLET SURVIVAL WITH RAT EMBRYONIC FIBROBLAST COCULTURE

Objective Islets transplantation of pancreatic islets has been demonstrated to be a viable alternative to exogenous insulin therapy for diabets mellitus. However, it requires that the islets must have a good function. This study was design to investigate how to retain best the islet function in transplantation as cocultured with EFCs. Methods The EFCs were isolated from SD rat fetus and islets were purfied from SD rats. The viability of the isolated islets was assessed by AO/PI double staining. The islets were cultured alone(group Ⅰ), coculture with EFCs(group Ⅱ) and EFCs only(group Ⅲ). Islet viablity was assessed by mitochondrial activity and insulin release assay. Results Morphology assay: Islets in group Ⅱ exhibited excellent morphology, with greater than 95% staining positive as detected by AO/PI. Insulin release assay showed that: There was significantly a higher simulation indece(SI) in group Ⅱ as compared with group Ⅰ(P<0.05) except the first day. Mitochondrial activity: A total of the absorbance of the islet cells by MTT in 3,5,7,14 day between group Ⅰ and group Ⅱ showed significant difference(P<0.05, or P<0.01).Conclusion This preliminary series of controlled experiments suggest that cocultrue of