CAJ | 학술논문

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.
목적 관찰대서장신경간세포(ENSCs)적배양방법,탐토재동충유문기이식적가행성.방법 종태령20d대서소화도제취ENSCs,체외배양、확증급감정;PKH-26표기후,주사이식지SD유서유문기층,분별재이식술후3d급1、2주절편관찰이식세포적활력급분포.결과 제취적세포경형태학화면역학감정위ENSCs,경류식세포의검측β-Ⅲ형미관단백(TUJ-1)양성세포수위12.3％,효질섬유산성단백(GFAP)위21.7％,소단백(Nestin)위10.8％,Ret위1.4％.이식후가이관찰도표기세포재유문기층보지정상적세포형태,수착시간연장,이식세포적분포종불규칙성전위규칙성배렬,병유향점막하층천이적적상.2주후,세포잉존활량호.결론 ENSCs이식후능재수체유문기층내존활、천이.
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,proliferated and labeled with PKH-26,then they were transplanted into the pylorus wall of rats by injection.Survival,morphology,and migration of ENSCs were observed at 3rd day,1st week,and 2nd week after transplantation.Results ENSCs were identified by morphology and immunology.Flow cytometry revealed that the percentage of β-Ⅲ tubulin (TUJ-1 positive ceils 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 positive 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 cells,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.