基于均匀设计优化延边黄牛卵母细胞体外成熟及核移植后卵裂体系

目的]应用均匀设计对延边黄牛卯母细胞体外成熟及核移植后的卵裂体系进行优化。方法]用抽吸法回收卵母细胞，在不同条件下进行卵母细胞的体外成熟培养，然后对成熟卵母细胞进行核移植、融合、激活以及胚胎的体外培养。采用均匀设计法，选用U10（10^3）均匀设计，比较了不同的卵巢保存温度、成熟培养时间以及卵泡直径大小对牛卵母细胞体外成熟率及卵裂率的影响。结果]延边黄牛卵母细胞体外成熟的最佳条件为：在卵巢保存温度为26℃或31℃的条件下，选取直径为8．0mm的卯母细胞成熟培养24h；卵母细胞核移植后卵裂的最佳条件为：在卯巢保存温度为26℃的条件下，选取直径为6．0mm或8．0mm的卵母细胞培养24h。结论]该结果对延边黄牛或其他种黄牛的育种及种群扩繁具有一定的参考意义。

Optimizing Yanbian Cow Oocytes Mature in vitro and Cleavage System after Nuclear Transfer Based on Uniform Design

Objective] The aim was to optimize Yanbian cow oocytes mature in vitro and cleavage system after nuclear transfer based on uniform design. Method] Oocytes were recovered by aspiration method, and oocytes were matured in vitro （IVM） with different conditions, and then carried out nucleus transfer, fusion, activation and in vitro culture （IVC） of embryo. Effects of ovary storage temperature, maturation time and follicular diameter size on in vitro maturation and cleavage rates of cow oocytes were compared. Result] The best conditions of IVM of Yanbian cow oocytes was that： the oocytes of 8 mm diameter were matured in vitro for 24 hours when the ovaries were stored at 26℃ or 31 ℃. The best cleave conditions after nucleus transfer of oocytes was that： the oocytes of 6 mm or 8 mm diameter were cultured in vitro for 24 hours when the ovaries were stored at 26℃. Conclusion] The result has some reference to Yanbian cow and other cow breeding and population expanding propagation.