摘要
目的：探讨一种分离纯化小鼠胰岛细胞的优良新型方法,经同种异体糖尿病小鼠肾被膜下胰岛移植建立动物模型.方法：（1）胰岛分离及检测：采用胆总管内胶原酶逆行灌注消化胰腺的方法分离胰岛,淋巴细胞分离液单一密度梯度离心法纯化胰岛.利用双硫腙对胰岛进行特异性染色来计算胰岛产量及纯度,台盼兰染色鉴定胰岛细胞的活性,以葡萄糖刺激胰岛素释放来检测胰岛功能.（2）胰岛移植：空白对照组（n =12）,糖尿病鼠组（n=12）,对糖尿病鼠组行同种异体鼠肾被膜下胰岛移植,观测血糖及生命体征变化.结果：每只小鼠可获得150～200个高质量胰岛,纯度及活性均高于90 ％,葡萄糖刺激后胰岛素释放量明显增加（SI=3）.糖尿病小鼠经胰岛移植后,1～3 d内,血糖均降至11.1 mmol/L以下.结论：采用新改进的分离纯化胰岛方法,可获得高质量、高功能的胰岛细胞.经同种异体糖尿病小鼠肾被膜下胰岛移植动物模型的成功建立,为进行人胰岛小鼠移植及人胰岛同种异体移植提供重要试验价值.
Objective：To discuss an excellent new method for isolating and purifying islet cells of mice,and to establish animal model through islet transplantation under the renal capsule of mice with allogeneic diabetes.Methods：（1） Pancreas islet isolation and detection：pancreas islet was isolated by perfusing the collagenase retrograde into the digestive pancreas in common bile duct,and purified with single density gradient centrifugation of lymphocyte separating medium.Specific staining was conducted for the pancreas with dithizone to calculate the output and purification of pancreas islet.The activity of islet cells was identified with trypan blue staining,and the functions of pancreas islet were detected through stimulating the insulin release with glucose.（2） Pancreas islet transplantation：blank control group （n =12）,mice with diabetes （n =12）,islet transplantation was carried out under the renal capsule of mice with allogeneic diabetes,for observing the variations in blood sugar and vital signs.Results：High-quality pancreas islets （150 ～ 200） could be obtained from each mouse,and the purity and activity were above 90％.The insulin release amount after the stimulation from glucose increased obviously （SI=3）.After the islet transplantation,the blood sugar of mice with diabetes decreased below 11.1 mmol/L within one to three days.Conclusion：With the newly-improved approach for isolating and purifying the pancreas islet,high-quality and highfunction islet ceils can be obtained.The successful establishment of the animal model for islet transplantation under renal capsule of rats with allogeneic diabetes can provide significant test value for the islet transplantation and allogeneic transplantation for human.
出处
《广西医科大学学报》
CAS
2014年第1期27-30,共4页
Journal of Guangxi Medical University
基金
教育部留学回国人员科研启动基金项目[No.教外司留(2011)1568号]
广西研究生教育创新计划资助项目(No.YCSZ2012040)
广西自然科学基金资助项目(No.2011GXNSFC018023)
广西医学科学实验中心开放基金专项项目(No.KFJJ2011-28)
(No.桂卫Z2010348)
关键词
淋巴细胞分离液
胰岛移植
小鼠
动物模型
lymphocytes separation
islet transplantation
mice
animal model
