摘要
目的探寻提高大鼠胰岛细胞分离纯化技术的方法。方法通过用胶原酶灌注及不连续密度梯度离心法分离纯化胰岛细胞,用DTZ染色计算胰岛细胞的纯度;通过体外葡萄糖刺激胰岛素分泌试验和糖尿病大鼠移植判定胰岛细胞功能。结果纯化后每只胰腺获得(768±135)个胰岛细胞,纯度为(81.5±11.6)%;纯化后细胞形态完好,活度大于92%。体外葡萄糖刺激胰岛素分泌实验,低糖情况下胰岛素分泌量为(23.16±4.13)mU/L,高糖情况下为(36.82±4.34)mU/L,两者有显著性差异(P<0.05)。胰岛移植后,实验组48 h后血糖值降至正常。结论用胰管内胶原酶灌注及不连续密度梯度法纯化可获得较高纯度、功能均良好的胰岛细胞。
Objective To explore a good methods for isolation and purification of rat islets.Methods The islets were isolated and purificated by collagenase perfusion and discontinuous gravity gradient centrifugation method,and counted by dithizone staining.The standard of isolated live cells were justified by trypan blue staining.The function of purified islets was determined by glucose stimulated insulin release test and transplantation of rat with streptozocin-induced diabetes.Results（768±135） islets were recovered after purification.The average purity was（81.5±11.6）%,the viability of islets was more than 92%.When inspected by glucose stimulation,the secreted insulin concentration was（23.16±4.13）mU/L when stimulated by low concentration glucose and（36.82±4.34）mU/L by high concentration glucose.There was significant difference between the two phases（P〈0.05）.The concentration of blood sugar recovered to normal level after two days in the group of islet transplantation.Conclusion Good morphology and function islet cells are got through collagenase perfusion and centrifugation on gradients method.
出处
《实用临床医学（江西）》
CAS
2008年第12期12-14,共3页
Practical Clinical Medicine
关键词
胰岛
分离
纯化
动物
实验
大鼠
islets
isolation
purification
animals,laboratory
rats
