不同蔗糖浓度、供体细胞保存温度和融合温度对小鼠体细胞克隆胚胎发育的影响  

Effects of different sucrose concentrations,conserved temperatures for donor cells and electro-fusion temperatures on the development of somatic cell nuclear transfer embryo in mouse

作　　者：彭礼繁[1,2] 罗光彬[1] 白文林[1] 闻福安[1] 任建诚[1] 冷义福[1] 

机构地区：[1]沈阳农业大学动物胚胎工程实验室,辽宁沈阳110161 [2]深圳武警医院生殖中心,广东深圳518023

出　　处：《广东畜牧兽医科技》2011年第3期26-30,共5页Guangdong Journal of Animal and Veterinary Science

摘　　要：以昆明小鼠为实验动物,经过超数排卵后得到卵丘卵母细胞复合体(COCs),以卵丘细胞为核供体细胞,卵母细胞为核受体细胞进行核移植,分析比较了不同蔗糖浓度对小鼠卵母细胞去核时间、去核率及重构胚体外发育的影响,并探讨了供体细胞的保存温度和融合温度对重构胚体外发育的影响。结果表明,各蔗糖浓度组的去核率均达到100%,但3%、4%、5%蔗糖浓度组的平均显微操作时间明显短于0%、1%、2%蔗糖浓度组,而4%和5%蔗糖浓度组的重构胚发育率均显著低于1%、2%、3%蔗糖浓度组;卵丘细胞保存在室温组的细胞融合率和体外发育率均显著高于高温组、低温组及温度变化组;25℃组细胞融合率显著高于37℃组。由此可见,显微操作液中添加3%蔗糖不仅使卵子的核明显,而且大大缩短卵子在体外操作时间,有利于重构胚的体外发育。卵丘细胞的室温保存和室温融合温度更有利于重构胚的体外发育。KunMing mouse were used as experiment animals in this study.Cumulus-oocyte complexes were obtained after superovulation.Cumulus cells were used as donor cells and oocytes were used as recipient cells.Mouse reconstructed embryo were obtained by somatic cell nuclear transfer.The effect of the different sucrose concentrations on in vitro enucleating time,in vitro enucleating rate of mouse oocyte and in vitro development of mouse reconstructed embryo,the effect of the different conserved temperatures for donor cells and electro-fusion temperatures on the development of reconstructed embryo were analyzed and compared.The results showed that enucleate rates of different sucrose concentrations in micromanipulation medium were all 100%,the average micromanipulation time in this groups of 3%,4%,5% sucrose concentrations were more shorter than that of 0%,1%,2% sucrose concentrations groups.But in vitro development rate of reconstructed embryo in micromanipulation medium with 4%,5% sucrose concentrations were significantly lower than that of 1%,2%,3% sucrose concentrations groups.Fusion rate and in vitro development rate in the room temperatures were significantly higher than that in warm temperature groups,cool temperature groups and changing-temperature groups.Fusion rate at 25℃ were significantly higher than that at 37℃.In Conclusion,3% concentration sucrose was the best to manifest DNA nuclear zone in the cytoplasm,shorten in vitro oocyte enucleate time and make enucleated operation quick for the in vitro development of reconstructed embryo.Room temperature for conservation of cumulus cells and fusion was good for in vitro development of reconstructed embryo.

关 键 词：小鼠 蔗糖浓度 保存温度 融合温度 体细胞核移植 

分 类 号：S814.6[农业科学—畜牧学]