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。结果该组研究共分离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(74.70±7.84)%,活度平均为(94.10±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。
ObjectiveTo investigate the methods of islet isolation and purification so as to get more islet cells with high purity and good function for clinical transplantation.MethodsHuman pancreas was digested with Liberase HI.The islets were isolated by the improved Recordi＇s automated islet isolation system,followed by purification in a Ficoll continuous gradient centrifugation using Cobe 2991 cell separator.After purification,the islet cell yield and purity were evaluated by DTZ staining under microscope and the activity was evaluated by AO-PI staining under fluorescence microscope.The islet＇s function was assessed by insulin release assay in vitro.ResultsThe islets were isolated from thirty pancreases.After purification,an average of（316 626±191 972） islet equivalents（IEQ） was obtained from each pancreas,with an average purity of（74.70±7.84）% and activity of（94.10±2.19）%.The islets responded well to 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.ConclusionThe improved Recordi＇s automated islet isolation system can provide high-purity islet cells for clinical transplantation.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2010年第17期2626-2629,共4页
China Journal of Modern Medicine
基金
国家自然科学基金(No:30571759)
全军"十五"重大课题(No:04Z007)
福建省科技计划重大项目(No:2009Y4001)
福建省科技创新平台建设计划(No:2008J1006)
关键词
胰岛细胞
细胞分离
纯化
islet of Langerhans
cell isolation
cell purification
