水牛卵母细胞体外受精与早期胚胎发育的研究

本实验通过 3种水牛卵母细胞体外成熟培养系统的比较 ,与早期胚胎发育研究结果表明 ,3组是最好的培养系统 ,卵母细胞成熟率为 5 1.9% (2 8 5 4 ) ;卵裂率为 5 2 .76 %± 9.0 8% (193 35 9) ;囊胚率 (占授精卵数 )为 2 7.2 2 %± 9.17% (10 1 35 9)、(占分裂卵数 )为 5 2 .13%± 16 .32 % (10 1 193) ;孵化囊胚率为 83.34%± 11.5 6 % (85 10 1)。该系统为含 10 %FBS的TCM - 199液中添加促性腺激素FSH、LH ,类固醇激素E2 ,表皮生长因子 (EGF)和硫醇类化合物β—巯基乙醇 (β—ME)的较完善系统。据 39批次早期胚胎体外发育至囊胚 (343枚 )所需时间 (以授精当日为 0d计算 )分析 ,5d囊胚 2枚 (0 .6 % ) ;6d囊胚 89枚 (2 5 .9% ) ;7d囊胚14 9枚 (43.4 % ) ;8d囊胚 73枚 (2 1.3% ) ;9d囊胚 2 5枚 (7.3% ) ;10d囊胚 5枚 (1.5 % )。2头经超数排卵处理的母水牛 ,授精后第5d非手术回收 3枚囊胚 ,说明受精卵在体外发育速度比在体内发育慢 ,推测试管水牛胚胎移植的适宜时间 ,应是受体母牛站立发情后第 4～ 6d

Studies on in vitro Fertilization and Early Embryonic Development in Buffalo Oocytes

This study compared the efficiency of three kinds of in vitro maturation culture systems and investigated early embryonic development in buffalo oocytes.The data show that group 3 is the best culture system,with maturation rate of 51.9%(28/54);cleavage rate of 52.76%± 9.08%(193/359); the proportions of blastocyst from inseminated oocytes and from fertilized oocytes were 27.22%±9.17%(101/359)and 52.13 ±16.32%(101/193),respectively,the hatched blastocyst rate reached 83.34%±11.56%(85/101).Supplementation of TCM 199 with 10% FBS,gonadotrophin(FSH and LH),steroid hormone(E 2),epidermal growth factors(EGF)and thiol compound(β mercaptoethanol,β ME),employed for in vitro matruation in group 3 was optimal.According to analysis of the time that early embryos(n=343)developed up to blastocysts by 39 batches(inseminated day=day 0),2 blastocysts were observed on day 5(0.6%),89 blastocysts on day 6(25.9%),149 blastocysts on day 7(43.4%),73 blastocysts on day 8 (21.3%),25 blastocysts on day9(7.3%),5 blastocysts on day 10(1.5%),Superovulatory treatment was carried out in 2 buffalo cows,3 blastocysts were recovered by non surgical flushing on day 5 post insemination.The results indicate the developmental speed of fertilized buffalo oocytes in vitro is slower than those in vivo,we therefor gpeculate the optimal time that the embryos transfer to recipients for producing tube buffaloes is from day 4 to day 6 after heat.