猪输卵管上皮细胞对小鼠早期胚胎体外发育的影响  

Effect of swine oviduct epithelium cells on development of co-cultured mouse early embryos in vitro

作　　者：赵永贞[1] 刘西梅[2] 李莉[2] 乔宪凤[2] 魏雁[2] 曹斌云[1] 魏庆信[2] 郑新民[2] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]湖北省农科院生物技术研究所,湖北武汉430072

出　　处：《西北农林科技大学学报（自然科学版）》2007年第5期9-13,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"计划项目(2001AA216071)

摘　　要：研究了不同生殖时期猪输卵管上皮细胞对小鼠早期胚胎体外发育的影响,以此建立了适于早期小鼠转基因胚胎体外培养的猪输卵管上皮细胞共培养体系,并用PCR法对体外培养发育到不同阶段的小鼠转基因胚胎进行了外源基因整合检测。结果表明,在受孕初期的猪输卵管上皮细胞(无论是原代还是传代细胞)上,培养的小鼠1-细胞期胚胎的囊胚率显著高于发情间期的猪输卵管上皮细胞(P<0.01);在相同生殖时期猪输卵管原代和传代上皮细胞上,培养的小鼠1-细胞期胚胎的囊胚率差异不显著;在转基因胚胎不同发育阶段,PCR检测呈阳性的胚胎比率分别为:2-细胞期为96%,4-细胞期为63%,8-细胞期为43%,囊胚期为19%。由此可见,受孕初期的猪输卵管上皮细胞可促进与之体外共培养的小鼠早期胚胎的发育;随着早期小鼠转基因胚胎的发育,PCR检测呈阳性的转基因胚胎的比率逐渐降低。By comparing the effect of pig oviduct epithelium cells derived from different estrous cycle for culturing early transgenic mouse embryos,an effective co-culture system for culture of early embryos in vitro was built. The retention of foreign gene in mouse embryos was detected by PCR further. The results showed that there was no significant difference between the blastocyst rate of embryos co-cultured with pig oviductal epithelium cells （whether or not primary and passage cells） derived from pig oviduct during same estrous cycle. Blastocyst rate of embryos co-cultured with pig oviduct epithelium cells from pig oviduct during diestrus was significantly lower than metestrous. The rate of transgenic positive embryos in 2-cell, 4- cell,8-cell and blastocyst stage was 96% ,63% ,43% ,and 19% ,respectively. All in all,pig oviductal epithelium cells derived from pig oviduct during metestrous can improve the development of mouse early embryos when mouse early embryos were co-cultured with it. With development of early embryos microinjected with foreign gene,the rate of transgenic positive embryos decreased gradually.

关 键 词：显微注射 共培养 小鼠胚胎 转基因 

分 类 号：Q813.11[生物学—生物工程]