高压氧微重力培养提高冻存胰岛质量和移植效果的实验研究  

An experimental study on improving quality of murine cryopreserved islets cultured with hyperbaric RCCS and transplantation

作　　者：周毅[1] 刘锐[1] 吴舰宇[1] 代文杰[1] 宋春芳[1] 

机构地区：[1]哈尔滨医科大学第一临床医学院普外一科卫生部细胞移植重点实验室,黑龙江哈尔滨150001

出　　处：《中华普通外科杂志》2009年第3期221-224,共4页Chinese Journal of General Surgery

基　　金：黑龙江省青年基金项目（QC06C079）

摘　　要：目的探讨胰岛冻存前后经高压氧细胞旋转培养系统（HORCCS）培养后移植入糖尿病大鼠能否提高胰岛移植的效果。方法将分离纯化的大鼠胰岛分为：A.体外实验组：将各组大鼠胰岛经HORCCS培养或普通培养30d，检测细胞内DNA和胰岛素含量，胰岛存活率，胰岛素分泌水平。B.胰岛移植实验组：将各组大鼠胰岛经HORCCS培养或普通培养7d，然后移植，观察移植受体血糖和胰岛素水平。电镜观察各胰岛移植实验组中培养7d时的胰岛的超微结构改变。结果经高压氧RCCS培养14d的胰岛存活率及胰岛素分泌水平高于普通培养组（P〈0.05）。移植了经HORCCS培养的胰岛后，受体血糖在移植后2周即恢复为正常值，并维持到移植后10周。全部受体维持正常血糖耐受曲线。电镜下可见经HORCCS培养后，冻存复苏胰岛表面形成微小的孔道。结论胰岛冻存前后经高压氧RCCS培养后可以建立营养输送管道，不但有利于氧和营养物质的运输，更有利于胰岛内细胞的均匀一致的冻存，从而减少冻存对胰岛的损害，提高胰岛的分泌活性和成活率。Objective To investigate the therapeutic effect of frozen-thawed murine islets which were transplanted into diabetic rats after cultured with hyperbaric oxygenated rotary cell culture system （HORCCS）. Methods The purified rat islets were divided into two groups： A. In vitro experiment groups（IvEG） ： The rat islets in each subgroup were cuhured in HORCCS or common medium for 30 days, then evaluated for the intracellular DNA and insulin contents of islets, and the viability and insulin secreting level of islets. B. Islet transplantation experimental groups（TxEG）： The frozen-thawed islets were cultured in HORCCS or common medium for 7 days, and then transplanted into the recipients. We observed the blood glucose level （BGL） and insulin secreting level in the recipients as well as the uhrastructure change of islets in TxEG. Results The viability and insulin secreting level of islets cultured with HORCCS at 14^th day were much higher than those cultured with common medium （P〈0.05）. The blood glucose level in recipients transplanted with islets cultured with HORCCS recovered to normal value at the 2nd week and lasted for 8 weeks. All these recipients maintained the normal glucose tolerance curve. Electronic microscopy found microchannel outlets on the surface of the frozen-thawed islets cultured with HORCCS. Conclusions Frozen-thawed islets cultured with HORCCS could establish nutrient transmission microchannels, which were not only capable of oxygen and nutrients transmission, but also improving cryopreservation solution to diffuse inside the islet ceils evenly and uniformly. So this method not only lessens islet damage from cryopreservation, but also improves the effect of transplantation.

关 键 词：胰岛移植 高压氧 大鼠 WISTAR 冻存 微重力 旋转培养 

分 类 号：R686[医药卫生—骨科学]