成人胰岛细胞分离纯化技术  

Modified method of human islet isolation and purification for clinical transplantation

作　　者：陈津[1] 黄梁浒[1] 王庆华[1] 马予洁[1] 杨顺良[1] 蔡锦全[1] 谭建明[1] 

机构地区：[1]南京军区福州总医院全军器官移植中心,福州350025

出　　处：《中华细胞与干细胞移植（电子版）》2009年第1期35-37,共3页

基　　金：国家自然科学基金（30571759）; 全军＂十五＂重大课题（04Z007）; 福建省科技厅重大攻关课题（2002Y007）

摘　　要：目的探讨成人胰岛细胞分离纯化技术,为胰岛细胞移植治疗1型糖尿病提供大量高质量的胰岛细胞。方法使用LiberaseHI复合胶原酶和改良的Recordi自动胰岛细胞分离技术分离胰岛,采用COBE2991细胞淘洗仪连续密度梯度离心纯化胰岛细胞,用DTZ和AO-PI染色显微镜下评价胰岛细胞的数量、纯度和活性。结果本组研究共分离30例胰腺的胰岛,获得胰岛细胞数量平均为（316626±191972）IEQ,平均每克胰腺收获胰岛细胞3389IEQ,收获的胰岛纯度平均为（74.7±7.84）﹪,活度平均为（94.1±2.19）﹪,胰岛细胞刺激指数平均为3.68±0.58。在此基础上,成功实施7人12次的成人胰岛细胞移植。结论成功建立了改良的Recordi自动胰岛分离技术,获得大量高纯度有活性的胰岛细胞,为开展胰岛细胞临床移植提供了保障。Objective To investigate the methods of islet isolation and purification to get more islet cells with high purity and good function for clinical transplantation. Methods Human pancreas was digested with Liberase HI.The improved Recordi’s automated islet isolation system was used,followed by purification in a Ficoll continuous gradient centrifugation using Cobe 2991 cell separator.After purification,the islet cell yield and purity was evaluated with microscope by DTZ staining,and the activity was evaluated with fluorescence microscope by AO-PI staining.The islet’s function was assessed by insulin release assay in vitro.Results Thirty pancreases were isolated.After purification,an average of（316626±191972）islet equivalents（IEQ）was obtained from each pancreas,with an average purity of（74.7±7.84）﹪and viabiling of（94.1±2.19）﹪The islets responded wellto glucose stimulation with a 3.68-fold increase of insulin secretion over the basal level. Seven patients with type 1 diabetes accepted islet transplantation with expected outcomes. Conclusion The improved Recordi’s automated islet isolation system can provide more high-purity islet cells for clinical transplantation.

关 键 词：胰岛细胞 细胞分离 纯化 

分 类 号：R283[医药卫生—中药学]