大鼠胰岛复合猪小肠黏膜下层体外构建组织工程胰腺的实验研究  

Artificial pancreas composing of rat islets and swine small intestinal submucosa construction

作　　者：宋勇[1] 薛武军[1] 田普训[1] 丁小明[1] 冯新顺[1] 田晓辉[1] 宋焕瑾[1] 李杨[1] 

机构地区：[1]西安交通大学医学院附属第一医院肾病中心,西安710061

出　　处：《中华细胞与干细胞杂志（电子版）》2012年第2期19-23,共5页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：陕西省"13115"科技创新工程重大科技专项项目计划(2007ZDKG-67);国家自然科学基金(81270548);卫生部临床学科重点项目(胰岛移植物营养支持及免疫保护的研究);陕西省自然科学基金项目(2007C212);西安交大医学院附属第一医院基金青年创新项目(2006YK4);陕西卫生厅科研基金(08D25)

摘　　要：目的探讨用猪小肠黏膜下层(SIS)作支架,复合大鼠胰岛体外构建组织工程胰腺的可行性。方法用改良的Ⅴ型胶原酶消化法方法分离纯化大鼠胰岛并培养,再与SIS进行体外复合培养。然后用相差显微镜、透射电镜和扫描电镜检查胰岛的形态,用丫啶橙/碘化丙啶(AO/PI)双染法观察胰岛的生长及细胞活性,用胰岛素释放试验评价胰岛细胞的分泌情况。采用独立t检验,分别比较在高糖、低糖条件下,SIS组与对照组胰岛素释放的差异有无统计学意义。结果胰岛在SIS材料上黏附、生长,细胞功能活跃,AO/PI双染法显示平均存活率92﹪,高糖下胰岛素释放量为(37.79±4.27)mmol/L,明显高于对照组[(28.69±3.58)mmol/L,t=12.678,P=0.000]。胰岛在SIS上成复层生长,复合培养时间明显延长。结论 SIS与胰岛在体外复合培养可以构建出类似生物膜的组织工程胰腺,为进一步研究体内胰岛移植奠定基础。Objective To investigate the feasibility of artificial pancreas construction with islets in a tissue scaffold of small intestinal submucosa（SIS）.Methods Pancreatic islets were isolated and purified using a modified method with collagenase.The islets co-cultivated with SIS were observed by phase contrast microscope,electronic microscope and scanning electronic microscope.Islet growth and secretion function were evaluated by AO/PI staining and high-glucose stimulating test.Results The islets adhered to and grew on SIS,with active cell function.High-glucose stimulating insulin secretion was（37.79 ± 4.27） mmol/L in SIS group,significantly higher than that of control group [（28.69 ± 3.58） mmol/L,t = 12.678,P = 0.000].Islets grew well in layers,with a 92﹪ viability,and the survival time was significantly longer.Conclusion Coculture of SIS and islet in vitro can construct an artificial pancreas similar to the biomembrane,thus providing a base for further study of islet transplantation in vivo by tissue engineering technique.

关 键 词：胰岛 小肠黏膜下层 组织工程 

分 类 号：R587.1[医药卫生—内分泌]