中国实验动物学报2018,Vol.26Issue(1):45-51,7.DOI:10.3969/j.issn.1005-4847.2018.01.007
NOD小鼠胰岛的分离及其在体内外的生物学特性
Isolation and characterization in vitro and in vivo of pancreatic islets from NOD mice
摘要
Abstract
Objective This study was conducted to establish a stable and highly efficient method for isolation and purification of pancreatic islets from NOD mice and to evaluate their characteristics in vitro and in vivo. Methods The is-lets were isolated from mouse pancreas using modified collagenase digestion and Ficoll density gradient centrifugation. The endocrine secretory function was assessed by insulin secretion in either low or high dose glucose stimulation. To evaluate the function of the graft,body weight and blood glucose were monitored,and IVGTT was performed. In addition,to assess sur-vival of the implanted islets,Pathology using HE staining and insulin immunostaining of the graft were performed. Results The average islet yield was 116 ± 12 islets/pancreas and purity was higher than 90%. Compared with islets from Kunming mice,the islets isolated from NOD mice were poorly responsive to glucose challenge. Blood glucose levels and body weight changes of the islet-transplanted diabetic mice were significantly improved compared with the sham-operated mice. In addi-tion,blood glucose levels in vivo after an IVGTT also significantly improved. However,these improvements were only main-tained for 2 weeks. Furthermore,HE staining and immunostaining assays demonstrated that there were insulin-positive cell clusters and lymphocyte infiltration in the graft-bearing kidney. Conclusions A large number of quality islets can be isola-ted and purified from NOD mice by using the modified mouse islet isolation method, which can be used to develop thera-peutic strategies to protect transplanted islets from rejection and autoimmune attack.关键词
非肥胖性糖尿病/胰岛/分离纯化/移植/小鼠Key words
NOD/pancreatic islets/isolation and purification/transplantation/mice分类
生物科学引用本文复制引用
舒冠男,滕夏虹,许倩倩,邹春林..NOD小鼠胰岛的分离及其在体内外的生物学特性[J].中国实验动物学报,2018,26(1):45-51,7.基金项目
国家自然科学基金项目(No.81360123),广西自然科学基金项目(No.2014GXNSFDA118030).Funded by National Natural Science Foundation of China(No.81360123)and Guangxi Natural Science Foundation(No.2014GXNSFDA118030). (No.81360123)