摘要
利用活体取卵(OPU)所得的卵母细胞与牛耳成纤维细胞构建牛体细胞核移植重构胚,并将所得的重构胚进行ACM(培养液)与不同滋养层细胞的共培养,分析不同滋养层细胞类型、传代次数、冻融处理对核移植胚胎体外发育的影响,探讨胚胎体外共培养中滋养层细胞的合适条件。结果表明:胎鼠成纤维细胞(MEF)培养组的囊胚率和胚胎细胞数(38.2%和135±2.1)显著高于牛颗粒细胞(GC)组(9.1%和101±4.6,P<0.05)和牛耳成纤维细胞(BAF)组(25.9%和115±3.1,P<0.05)。2～4代MEF细胞与胚胎共培养的效果(38.2%和135±2.1)明显好于原代MEF细胞(24.7%和108±3.1,P<0.05)或5～8代MEF细胞(22.8%和102±2.8,P<0.05),新鲜的MEF细胞比冻融处理后的MEF细胞培养效果好(38.2%vs.27.7%,135±2.1vs.110±1.7,P<0.05)。因此,用新鲜的2～4代MEF细胞作为共培养体系中的滋养层细胞有利于牛核移植胚胎的早期发育。
Cloned embryos constructed by bovine ovum pick up （OPU） oocytes and bovine adult fibroblast cells were co-cultured with different feeder cells in ACM （culture media）. Effects of co-culture cell types, passages and cryopreservation on the in vitro development of bovine cloning embryos were investigated. The result showed that the mouse embryonic fibroblasts （MEF） group achieved significantly higher blastocyst formation rate （38.2%） and the cell number of embryo （135±2.1） compared with granulosa cells （GC） group （9.1% and 101±4.6, P〈0.05） and bovine adult fibroblasts （BAF） group （22.8% and 102±2.8, P〈0.05）. Passages of co-culture cells sigoificantly affected tile blastocyst rate and the cell number of embryo, and MEF cells passaged to 2～4 were better than primary and 5～8 passages for co-culture （38.2% vs. 24.7% and 22.8%; 135±2.1 vs. 108±3.1 and 102±2.8, P〈0.05）; Cryopreservation of MEF cells went against the blastocyst development （27.7% vs. 38.2%, 110±1.7 vs. 135±2.1 ,P〈0.05）. In conclusion, co-culture with fresh 2～4 passages mouse embryonic fibroblasts could improve the development of bovine nuclear transfer embryos in vitro.
出处
《上海交通大学学报（农业科学版）》
2007年第6期519-524,共6页
Journal of Shanghai Jiaotong University(Agricultural Science)
基金
国家"863"计划基金资助项目(2005AA206140)
上海市科委重大攻关资助项目(05DZ19322)
关键词
胎鼠成纤维细胞
共培养体系
牛
体细胞核移植
mouse embryonic fibroblast
co-culture system
bovine
somatic cell nuclear transfer （SCNT）
