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不同因素对牛卵母细胞胞质内精子注射效果的影响
引用本文:原巨强,陈静波,刘赛,海里且木,赵晓娥,马保华.不同因素对牛卵母细胞胞质内精子注射效果的影响[J].西北农业学报,2007,16(6):8-13.
作者姓名:原巨强  陈静波  刘赛  海里且木  赵晓娥  马保华
作者单位:1. 西北农林科技大学动物科技学院,农业部家畜生殖内分泌与胚胎工程重点开放实验室,陕西杨凌,712100;新疆畜牧科学院,新疆乌鲁木齐,830000
2. 新疆畜牧科学院,新疆乌鲁木齐,830000
3. 西北农林科技大学动物科技学院,农业部家畜生殖内分泌与胚胎工程重点开放实验室,陕西杨凌,712100
基金项目:陕西省重大科技专项计划;西北农林科技大学校科研和教改项目
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。

关 键 词:显微受精  卵母细胞体外成熟  精子状态  聚乙烯吡咯烷酮  ICSI卵体外培养  
文章编号:1004-1389(2007)06-0008-06
收稿时间:2007-03-26
修稿时间:2007-06-10

Influence of Different Factors on Intractoplasmic Sperm Injection in Bovine Oocytes
YUAN Ju-qiang,CHEN Jing-bo,LIU Sai,HAI Li-qie-mu,ZHAO Xiao-e and MA Bao-hua.Influence of Different Factors on Intractoplasmic Sperm Injection in Bovine Oocytes[J].Acta Agriculturae Boreali-occidentalis Sinica,2007,16(6):8-13.
Authors:YUAN Ju-qiang  CHEN Jing-bo  LIU Sai  HAI Li-qie-mu  ZHAO Xiao-e and MA Bao-hua
Institution:Key Laboratory of Animal Reproductive Endocrinology and Embryo Biotechnology, Ministry of Agriculture, College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China;Xinjiang Academy of Animal Science, Urumqi Xinjiang 830000, China;Xinjiang Academy of Animal Science, Urumqi Xinjiang 830000, China;Key Laboratory of Animal Reproductive Endocrinology and Embryo Biotechnology, Ministry of Agriculture, College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China;Xinjiang Academy of Animal Science, Urumqi Xinjiang 830000, China;Key Laboratory of Animal Reproductive Endocrinology and Embryo Biotechnology, Ministry of Agriculture, College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China;Key Laboratory of Animal Reproductive Endocrinology and Embryo Biotechnology, Ministry of Agriculture, College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China
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.
Keywords:Microfertilization  In vitro maturation of oocyte  Sperm state  Polyvinylpyrrolidone  In vitro culture of the ICSI zygotes  Bovine
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