肠神经干细胞幽门移植的研究  

Transplantation of enteric neural stem cells into pylorus of rats

作　　者：朱利斌[1] 李仲荣[1] 赵艳民[1] 刘征吉[1] 李东[2] 

机构地区：[1]温州医学院附属育英儿童医院外科,325027 [2]温州医学院生命科学院

出　　处：《中华实验外科杂志》2012年第12期2491-2493,共3页Chinese Journal of Experimental Surgery

基　　金：浙江省卫生厅资助项目（2009A149）;温州市科技局资助项目（Y20070064）;浙江省自然科学基金资助项目（Y207272）

摘　　要：目的观察大鼠肠神经干细胞（ENSCs）的培养方法，探讨在同种幽门肌移植的可行性。方法从胎龄20d大鼠消化道提取ENSCs，体外培养、扩增及鉴定；PKH-26标记后，注射移植至SD幼鼠幽门肌层，分别在移植术后3d及1、2周切片观察移植细胞的活力及分布。结果提取的细胞经形态学和免疫学鉴定为ENSCs，经流式细胞仪检测β-Ⅲ型微管蛋白（TUJ-1）阳性细胞数为12．3％，胶质纤维酸性蛋白（GFAP）为21．7％，巢蛋白（Nestin）为10．8％，Ret为1．4％。移植后可以观察到标记细胞在幽门肌层保持正常的细胞形态，随着时间延．长，移植细胞的分布从不规则性转为规则性排列，并有向黏膜下层迁移的迹象。2周后，细胞仍存活良好。结论ENSCs移植后能在受体幽门肌层内存活、迁移。Objective To observe the in vitro culture of enteric neural stem cells （ENSCs） in rats, and explore the possibility of homotransplantion of ENSCs into the pylorus of rats. Methods ENSCs were obtained from the alimentary tract of the SD rats in embryonic age 20th day, cultured in vitro, prolifer- ated and labeled with PKH-26, then they were transplanted into the pylorus wall of rats by injection. Sur- vival, morphology, and migration of ENSCs were observed at 3rd day, 1st week, and 2nd week after trans- plantation. Results ENSCs were identified by morphology and immunology. Flow cytometry revealed that the percentage of ^-]]I tubulin （TUJ-1 positive cells in ENSCs was 12. 3%, that of glial fibrillary acidic protein （GFAP） positive cells was 21.7% , that of nestin positive cells was 10. 8% , and that of Ret posi- tive cells was 1.4%, respectively. Labeled ENSCs by PKH-26, a red fluorescence, maintained the normal morphology in pylorus walls. With time going on, the distribution of the grafted ceils, which were irregular primarily, became regular, and immigrated toward submucosa. Grafted cells were survival well within 2 weeks. Conclusion Transplanted ENSCs can survive and migrate in pylorus wall of rats.

关 键 词：肠神经干细胞 移植 先天性巨结肠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]