牛卵母细胞对成纤维细胞重编程作用  

The Bovine Oocyte Plays a Role in Reprogramming of Fibroblast Cell

作　　者：吴苏君[1] 马晓玲[1] 赵贵民[1] 杨文琳[1] 云彦[1] 雷安民[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西省干细胞工程技术研究中心,陕西省农业分子生物学重点实验室,杨凌712100

出　　处：《农业生物技术学报》2011年第4期719-724,共6页Journal of Agricultural Biotechnology

基　　金：国家高技术研究发展计划(863)项目(No.2008AA101010)资助

摘　　要：转录因子nanog和oct-4是细胞具有多潜能性的关键调控因子,一般认为它们的表达是细胞具有多潜能性和自我更新能力的标志。本研究将牛成纤维细胞(BEF422)分别移入GⅤ期和MⅡ期卵母细胞,体外培养24h后,RT-PCR检测其nanog和oct-4基因的表达情况,并用免疫荧光染色法对这两个基因在移植卵中的分布进行定位。结果表明,牛成纤维细胞移植入GⅤ期和MⅡ期卵母细胞培养后,nanog和oct-4基因均被诱导表达,并且明显定位于移植细胞的细胞核上,而成纤维细胞组和未注射成纤维细胞的空白卵母细胞组中均未检测到nanog和oct-4的表达。说明GⅤ期和MⅡ期卵母细胞均能激活成纤维细胞中转录因子nanog和oct-4的表达,使其发生重编程。本研究选取GⅤ期和MⅡ期这两个关键时期的卵母细胞为研究对象,为进一步解释卵母细胞中含有的与供体细胞重编程相关的物质及作用机理提供了一定的基础资料。Transcription factor nanog and oct-4 are the fundamental features of pluripotent Stem Cell,which are the key factors to keep their pluripotency and self-renew ability.We transplanted 10 fibroblast cells（BEF422） into bovine oocytes at GⅤ and MⅡstages respectively,and cultured the oocytes for 24 h in vitro,than we used the RT-PCR and immunofluorescence technique to check the expression and location of nanog and oct-4 in the injected oocytes.The results showed that nanog and oct-4 were all expressed in the injected oocytes and located on transplanted cells nuclear.However,they weren＇t detected in fibroblast cells and oocytes control group;This result demonstrated that GⅤ and MⅡ stage oocytes both can activate the transcription factor nanog and oct-4 expression to reprogram the fibroblast cells.This study will lay a foundation for further verifying the reprogramme mechanism and related material on donor cells and oocytes.

关 键 词：成纤维细胞 GV期/MⅡ期卵母细胞 重编程 NANOG OCT-4 

分 类 号：S823.3[农业科学—畜牧学]