摘要
目的 初步探索人胎胰岛细胞的分离培养方法，以期深入研究胰岛细胞的凋亡机制及开展人胰岛细胞移植。方法 采用机械分离、体外胶原酶消化，并通过组织块联合培养、转皿、时间消化差法获取人胎胰岛细胞，观察胰岛细胞生长及开展人胰岛素释放情况。结果 双硫腙（Dithizone）染色显示胰岛纯度约80％～90％，细胞培养至7～9d为其对数生长期，培养至11d可测到胰岛素释放高峰，培养至15d仍具有一定的生物活性。结论 机械分离消化、转皿、时间消化差法可获得生物活性较好的胰岛细胞，为深入探讨胰岛细胞凋亡、胰岛细胞移植提供细胞基础。
Objective To establish a method for isolation and culture of human embryo pancreatic islet cells for the further understanding of the apoptosis of the cells. Methods Human embryo pancreatic islet cells were isolated by the perfusion of collagenase after the manual separation, and were purified and harvested by the co-cultivation, time-lag digestion. The yield and viability were assessed by Dithizone dye test. The morphology alteration of culturing islet cells in vitro was observed, together with the concentrations of insulin in the supernation in different culturing time point. Results The average purification of pancreatic islet cells was 80%-90%. The logarithmic growth period was 7-9d of the cultivation. The releasing of the insulin got its ultimate at about lld, and biologic activity of the cultivated cell could be found in about 15d. Conclusion The method used shows satisfactory result in isolating and purifing the pancreatic islet cells,and the cultured islet cells grow and function well, proved by PCM and insulin releasing detection, which provides cell sources in the further study of cell death and pancreatic islet cell transplantation.
出处
《重庆医学》
CAS
CSCD
2006年第9期783-784,共2页
Chongqing medicine
关键词
糖尿病
胰岛细胞
移植
分离
培养
diabetes mellitus
islet cell
transplantation
