胰岛细胞与神经细胞和胶质细胞的共同培养研究

The coculture of pancreatic islets with neurons and gliacytes

目的 验证神经细胞和胶质细胞 (NG)对胰岛细胞的营养和生长促进作用。方法 采用SD大鼠NG与SD大鼠及新生猪胰岛样细胞团 (ICCs) ,进行同种属和异种属细胞间的共同培养。用倒置显微镜、透射和扫描电镜观察两种细胞的生长状况 ,MTT法测定细胞的活力 ,放免法和比色法测定胰岛素和淀粉酶的含量 ,免疫组化鉴定共同培养的细胞。结果 共同培养能显著延长同种属及异种属胰岛细胞的生存期 (>1月 ) ,增加ICCs的活力和胰岛素的分泌 (P <0 0 5 ) ,减少胰淀粉酶的分泌 (P <0 0 5 )。抑制成纤维细胞的过度增生。结论 共同培养能为胰岛细胞的生长提供一个优良的微环境。这为脑内胰岛移植提供了一个新依据 ;为胰岛细胞的体外保存提供了一个新方法。在移植前与NG共同培养以消除外分泌部细胞的影响 ,也是一个好策略。

Objective To identify whether the neurons and gliacytes(NG) have beneficial effects on the growth and survival of islets. Methods The ICCs of adult SD rats and newborn porcine were cocultured with the neurons and gliacytes of newborn SD rats.The growth appearnce of ICCs was inspected by inverted microscope and electricmicrocope.The viability of ICCs and the content of insulin and amylopsin were measured. The β-cells in the coculture were identified by immunohistochemistry.Results The survival span of ICCs in culture was prolonged significantly when cocultured with neurons and gliacytes. The insulin content and the viability of the ICCs were increased,the amylopsin content was decreased.Conclusion The neurons and gliacytes gave better microcircumstances for the ICCs survival and growth.This results should provide another evidence for the intracerebral islets transplantation.The coculture of ICCs with NG cells could provide a new method for long time preservation of the ICCs,and could inhibit fibroblast in vitro.