家兔胰岛的分离纯化研究

目的探讨家兔胰岛分离、纯化的方法及其计数和活性的测定。方法选用日本大耳自家兔。予V型胶原酶于38℃内水浴外消化。采用Ficoll 400不连续密度梯度离心法对胰岛进行纯化，并对获得的胰岛进行DTZ染色计数。同时计算胰岛当量（IEQ）数。予台盼蓝染色法分析胰岛活性。结果DTZ染色纯化前每只家兔胰腺收获胰岛细胞数为15046±2733。IEQ数为：4635±554；化后胰岛数量为：5300±609，IEQ数为：1213±218；胰岛细胞活率〉90％，胰岛纯度为〉60％。结论家兔胰腺消化分离后得到胰岛的数量和IEQ均较多，且动物来源广泛、实验成本较低，预示其在胰岛移植中可作为胰岛来源。

Method for isolation and purification of rabbit pancreatic islets

Objective To develop a method for isolation and purification of rabbit pancreatic islets. Methods Pancreatic islets were isolated from Japanese white rabbits,the islets were digestion by collagenase V at 38℃ ,discontinuous ficoll 400 density gradient centrifugation was used to purify the islets. Dithizone method was used for sample counting, islets equivalent quantity （IEQ）was calculated,the viability of islets was measured by trypan blue method. Results The digestion time of collagenase V was 35 minutes. Each pancreas harvested 15046±2733 islets and 4635±554 IEQ before purify,and those after purify were 5300±609 islets and 1213±218 IEQ. The purity was 〉60% and the viability rate of islets was 〉90%.Conclusion High quantity and IEQ islets can be harvested from rabbit pancreas by collagenase digestion, which can be used a source for islet transplantation.