体质量对大鼠胰岛分离纯化产量的影响  

Effect of body weight on the isolation,purification and yield of pancreatic islets in rats

作　　者：钱滔来[1] 王新华[1] 刘圣[1] 马亮[2] 任丹妮[2] 

机构地区：[1]上海市同济大学附属东方医院疼痛科,上海市200120 [2]上海市同济大学附属东方医院细胞治疗中心,上海市200120

出　　处：《中国组织工程研究与临床康复》2009年第44期8685-8689,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景：由于实验条件对胰岛的产量影响较大,因此需要在动物种属、动物周龄、体质量以及胶原酶的浓度、胰腺消化时间等方面进行优化组合,筛选出最佳条件,以期得到高产量的胰岛。目的：观察体质量因素对大鼠胰岛分离纯化产量的影响,筛选出获取高产量胰岛素的最佳体质量。设计、时间及地点：随机对照动物实验,于2008-01/12在上海市东方医院细胞治疗室及中心实验室等相关单位完成。材料：60只雄性SD大鼠按照体质量随机分为100-150g组、150-200g组、200-250g组、250-300g组、300g以上组,每组12只。方法：通过胆总管灌注胶原酶V来消化大鼠胰腺,分离胰岛,采用不连续密度梯度Ficoll离心法纯化胰岛。主要观察指标：以双硫腙染色鉴定胰岛,丫啶橙/碘丙啶染色鉴定胰岛细胞活率,根据胰岛直径计数所获胰岛的数量及胰岛当量,葡萄糖刺激胰岛素释放试验评估胰岛功能。结果：分离的大鼠胰岛具有较好的质量,胰岛纯度为95%左右,活率为90%左右,体外培养中生长良好。随着体质量增加,胰岛数量和胰岛当量明显增加（P〈0.05）,具有明显的相关性（r=0.918,r=0.922,P〈0.01）,并且最大直径的胰岛数量也在增加（P〈0.05）,以300g以上组所获得的胰岛数量及胰岛当量最多（P〈0.05）。在低糖和高糖刺激下胰岛素释放分别为（62.33±9.67）mU/L和（125.5±22.04）mU/L,差异具有显著性意义（P〈0.05）,刺激指数为2.01±0.21,提示胰岛细胞功能良好。结论：大鼠体质量是获得高产量胰岛的关键性因素之一,在其他条件不变的情况下,体质量高的大鼠所获得的胰岛产量较高。实验选择300g以上的大鼠进行胰岛分离纯化可获得稳定的高产量胰岛。BACKGROUND：As experimental condition has great influence on the yields of pancreatic islets,the best condition is screened by optimum combination of animal species,week old,body weight,concentration of collagenase,and digestion time of pancreas,in order to obtain the high yields of pancreatic islets.OBJECTIVE：To observe the effect of body weight on the yield of isolation and purification of pancreatic islets in rats,and to screen the best body weight to obtained the high yields of pancreatic islets.DESIGN,TIME AND SETTING：A randomized controlled animal experiment was performed in the Cell Therapy Center and the Central Laboratory of Shanghai East Hospital（Tongji University）from January to December in 2008.MATERIALS：Sixty male Sprague-Dawley rats were divided into five groups based on the body weight：100-150 g group,150-200 g group,200-250 g group,250-300 g group,and more than 300 g group,with 12 rats in each group.METHODS：Collagenase V was injected into the common bile duct of the rat for digestion of pancreas.The pancreatic islets was isolated and purified by using the discontinuous Ficoll density gradient centrifugation.MAIN OUTCOME MEASURES：The pancreatic islets were identified by dithizone staining,and the viability was assessed by acridine orange and propidium iodide staining.The yield of pancreatic islets and pancreatic islet equivalent was assessed based on the islets diameter.The islets function was measured by glucose-evoked insulin release.RESULTS：The isolated pancreatic islets had good quality,and grew well in vitro.The purity was about 95%,and the viability was about 90%.As the increasing body weight,the yield of pancreatic islets and pancreatic islet equivalent increased significantly（P〈0.05）,which showed that they had an obvious correlation（r=0.918,0.922,P〈0.01）.And the yield of islets and islet equivalent were highest in the more than 300 g group（P〈0.05）.The insulin release evoked by low and high concentration of glucose was（62.33±9.67）mU /L and（125.5±2

关 键 词：体质量 大鼠 胰岛 产量 

分 类 号：R318[医药卫生—生物医学工程]