【摘要】 采用胞质内精子注射法(ICSI)构建牛显微受精胚胎,探讨卵母细胞成熟培养时间、精子状态及精子操作液中聚乙烯吡咯烷酮(PVP)浓度对牛卵母细胞胞质内显微受精(ICSI)卵体外发育的影响。结果显示:成熟培养20 h、24 h和28 h的卵母细胞用于ICSI时,ICSI卵的形态完整率分别为94.6%(175/185)、97.2%(173/178)和96.4%(189/196)(P>0.05);成熟培养24 h和28 h的卵母细胞,其ICSI卵的卵裂率(73.4%和70.9%)和囊胚发育率(31.8%和29.6%)显著高于成熟培养20 h的卵母细胞(61.1%和20.6%)(P<0.05)。对成熟培养24 h的卵母细胞注射获能精子时,ICSI卵的卵裂率和囊胚发育率(72.7%和31.4%)高于未获能精子组(69.4%和29.3%)和不运动精子组(71.5%和30.4%),但无统计学差异(P>0.05)。采用含5%,10%和15%PVP的精子操作液处理获能精子后对成熟培养24 h的卵母细胞进行ICSI操作,PVP浓度为5%和10%时,ICSI卵的卵裂率(73.2%和66.9%)和囊胚发育率(32.7%和29.7%)差异不显著(P>0.05),但二者均显著高于15%PVP浓度组的卵裂率和囊胚发育率(51.0%和16.3%)(P<0.01)。结论认为,选用获能精子,并使用含5%PVP的精子操作液进行处理,对成熟培养24 h的牛卵母细胞进行ICSI操作,有利于ICSI卵的体外发育。更多还原

【Abstract】 This study investigated the effects of in vitro maturation time of oocyte,sperm state and the polyvinylpyrrolidone(PVP) concentration in the spermic manipulation medium on in vitro development of the intracytoplasmic sperm injection(ICSI) zygotes in bovine.The oocytes matured in vitro for 20,24 h and 28 h were used in ICSI,there was no effect of the maturation time of oocyte on the modality integrity rates of ICSI zygotes(94.6%,97.2%,96.4%,P> 0.05),the cleavage rates and blastocyst rates of ICSI zygotes were significantly higher with 24 h(73.4%,31.8%) and 28 h(70.9%,29.6% ) maturation than those with 20 h(61.1%,20.6%)(P< 0.05).Three different sperm states(capacitated sperm,without capacitated sperm and unmovable sperm) and the oocytes were matured in vitro for 24 h were used in ICSI,the higher cleavage and blastocyst rate of ICSI zygotes were obtained from the capacitated s1perm(72.7%,31.4%) than those from the without capacitated sperm and the unmovable sperm(69.4%,29.3% and 71.5%,30.4%),but there was no significant different(P>0.05).Three different PVP concentrations(5%,10% and 15%) in spermic manipulation medium were compared when the oocytes of in vitro maturation for 24 h and the capacitated sperm were used in ICSI,there was no different between 5% and 10% PVP concentration in the cleavage rates(73.2% vs.66.9%) and blastocyst rates(32.7% vs.29.7%,P>0.05) of ICSI zygotes,but the cleavage rates and blastocyst rates of ICSI zygotes were significantly higher than those with 15% PVP concentration(51.0 % and 16.3%)(P<0.01).The result indicated that the oocytes with in vitro maturation for 24 h and the capacitated sperm be used in ICSI,and the sperm was treated in the manipulation medium with 5% PVP,can improve the in vitro development of the ICSI zygotes in bovine.更多还原