大鼠胰岛的分离和纯化  

Isolation and Purification of the Islets of Langerhans in the Rats

作　　者：侯凌[1] 尹注增[2] 向莹[2] 李俊华[2] 陈实[2] 陈刚[2] 罗小平[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科,武汉430030 [2]华中科技大学同济医学院附属同济医院器官移植研究所,教育部/卫生部重点实验室,武汉430030

出　　处：《华中科技大学学报（医学版）》2009年第3期413-416,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家"十一五"科技支撑计划项目(No.2006BAI05A07);国家重点基础研究发展计划项目(No.2005CB522500)

摘　　要：目的探讨获得足够数量和较高纯度大鼠胰岛的分离和纯化方法。方法采用Ⅴ型胶原酶灌注消化法分离成年大鼠胰岛,Ficoll400非连续密度梯度法纯化胰岛。双硫腙(DTZ)染色鉴定培养的胰岛细胞,锥虫蓝染色检测细胞活性。葡萄糖刺激胰岛素释放试验评价胰岛细胞的功能。结果分离纯化后大鼠胰岛平均收获量为(480±30)IEQ,纯度大于90%,活性大于90%。在低糖和高糖刺激下,胰岛素分泌量分别为(12.28±0.89)mIU/L、(19.01±1.49)mIU/L,二者相比较,差异有统计学意义(P<0.05),刺激指数为(1.55±0.01)。结论胶原酶Ⅴ逆行灌注消化胰腺和Ficoll400非连续密度梯度纯化法,可获得高纯度高活率的大鼠胰岛。Objective To explore the method obtaining rat islets with enough quantity and higher purity. Methods Adult rat islets of langerhans were isolated by a collagenase V digestion technique and discontinuous Ficoll density gradient centrifugation method. Islets purity was assessed by dithizone staining. The viability of islets was assayed by trypan blue test. The function of purified islets was determined by glucose stimulated insulin releasing test. The changes of cultured islets were observed under a microscope. Results The total number of purified islets from one donor was （480±30） IEQ. The average purity and viability of islets were more than 90%, respectively. Insulin secretion of the islets was （19.01 ± 1.49） mIU/L under high glucose stimulation, which was higher than that under low glucose stimulation E（12.28±0.89） mIU/L, P〈0. 05]. The average insulin stimulation index was 1.55±0.01. Conclusion The digestion of collagenase V and discontinuous Ficoll density gradient centrifugation can obtain a high yield and high purity islets of pancreas in rats.

关 键 词：大鼠胰岛 分离 纯化 

分 类 号：R322.5[医药卫生—人体解剖和组织胚胎学]