套皿法在猪卵母细胞体外成熟培养中的应用

Application of nest dish culture method on in vitro maturation of porcine oocytes

试验表明,套皿无矿物油组卵母细胞成熟率与套皿矿物油组、单皿矿物油组差异不显著,单皿无矿物油组没有卵母细胞成熟;将各组成熟的卵母细胞分别进行体外受精,结果套皿无矿物油组卵母细胞的穿卵率、单精入卵率与套皿矿物油组、单皿矿物油组差异不显著,但雄原核形成率和卵裂率显著高于套皿矿物油组和单皿矿物油组(P<0.05)。套皿法有利于卵母细胞胞质成熟,促进胚胎生长发育,能够替代传统的单皿矿物油覆盖培养法。

To investigate the feasibility of nest dish without mineral oil culture method(nest dish method)on in vitro maturation(IVM)of porcine oocytes, we used nest dish or single dish. According to the using or no using of mineral oil, we divises the oocytes into four groups to culture:①nest dish without mineral oil, ②nest dish with mineral oil,③single dish without mineral oil,④single dish wit h mineral oil.The results showed no significant difference of maturation rate am ong group①?②and④.However, no matured oocytes were seen in group③.In order to study the effects of nest dish method on the subsequent development competence, we use the matured oocytes to fertilized in vitro.No significant effects can be seen on the percentage of sperm penetration, monospermy amog group①?②and ④.B ut the male pronuclear formation and cleavage rate of group①was significant hig her than group②and ④(P<0.05).Our study indicates that nest dish method can sub stitute for classical culture method covered with mineral oil, and it can also i mprove cytoplasm maturation and enhance embryo development competence.