大鼠肠神经干细胞同种消化道移植的实验研究

A primary study of homotransplantation of enteric neural stem ceils into the alimentary tract in rat

目的研究大鼠肠神经干细胞(ENSCs)在同种消化道壁内移植的可行性,为今后肠神经元发育缺陷性疾病的移植治疗提供依据。方法从胚胎SD大鼠消化道提取ENSCs,进行体外培养、扩增及PKH-26染料标记后,注射移植至幼SD大鼠结肠和幽门肌层,移植术后第3、7、14天观察移植细胞的形态、分布和分化情况。结果本研究提取的ENSCs,经PKH-26标记后能保持正常形态;荧光显微镜观察移植后不同时间结肠和幽门壁内均可见到红色荧光细胞.并保持正常细胞形态;随着移植后时间的延长,移植细胞的分布从不规则性转为规则性排列,并有向四周扩展迁移的迹象;移植术后14 d,可见部分移植的红色荧光细胞表达神经元标记物Tuj-1,提示向神经元分化的迹象。结论从大鼠消化道提取的ENSCs,移植入同种幼大鼠结肠和幽门肌层后,在宿主活体内能够存活,并有迁移和分化迹象。

Objective To explore the feasibility of homotransplantation of enteric neural stem cells （ENSCs） into the alimentary tract in rat, and to provide basis for the treatment of developmental defects of enteric nervous system. Methods ENSCs obtained from the alimentary tract of the embryonic SD rats, cultured in vitro, proliferated and labeled with PKH-26 were transplanted into colon and pylorus wall of young SD rat by injection. On the 3rd, 7th, 14th day after transplantation, morphology, differentiation and migration of ENSCs were observed. Results The study showed that ENSCs, labeled with PKH-26, morphologically remained normal. After transplantation, cells labeled with red fluorescent could be observed to maintain the normal morphology in the walls of colon and pylorus. With time prolonged, the distribution of the grafted cells, which was irregular when injected primarily, became regular. And there was evidence that they could disperse. On the 14th post-transplantation day, some grafted red fluorescent cell expression Tuj-1, a specific marker of neurons, suggested differentiation towards neurons. Conclusions ENSCs obtained from the alimentary tract of rat can survive the homotransplantation in the colon and pylorus of young rats, and they can migrate and differentiate.