| 绵羊成熟卵母细胞GMP玻璃化冷冻影响发育因素的研究 |
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| 引用本文： | 权富生,刘颖勋,刘利杰,沈钰,马会明,余源,张涌.绵羊成熟卵母细胞GMP玻璃化冷冻影响发育因素的研究[J].西北农业学报,2009,18(6):18-22. |
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| 作者姓名： | 权富生 刘颖勋 刘利杰 沈钰 马会明 余源 张涌 |
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| 作者单位： | 西北农林科技大学,动物医学院,动物生殖生理与胚胎工程农业部重点实验室,陕西杨凌,712100 |
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| 摘 要： | 利用GMP法(毛细玻璃管法)冷冻成熟的绵羊卵母细胞,旨在探讨冷冻和解冻处理方法对绵羊成熟卵母细胞发育潜力的影响.对比在玻璃化冷冻液中添加0.3 mol/L蔗糖;卵母细胞冷冻前用7.5 μg/ml CB预处理;卵母细胞去掉颗粒细胞冷冻;以及四步法解冻和三步法解冻对卵母细胞形态完整率、孤雌激活胚发育率的影响.结果显示:玻璃化冷冻液中添加蔗糖孤雌激活卵裂率达到56.70%,极显著高于对照25.40%(P<0.01),桑葚胚率(16.49%)比对照(4.76%)有明显提高(P<0.05);冷冻前用CB进行预处理,解冻后形态正常率为78.78%,明显高于对照61.11%(P<0.05),孤雌激活卵裂率(39.39%)也高于对照组(21.11%);去掉颗粒细胞裸卵与保留颗粒细胞成熟卵母细胞解冻后形态正常率、孤雌激活卵裂率及桑葚胚率、囊胚率之间无显著性差异(P>0.05);四步法解冻和三步法解冻后卵母细胞形态恢复及孤雌激活后的发育率,两组之间差异不显著(P>0.05).在玻璃化冷冻液中添加蔗糖及卵母细胞冷冻前用CB预处理有利于卵母细胞冷冻解冻后形态恢复及孤雌激活后的胚胎发育.
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| 关 键 词： | 玻璃化冷冻 成熟卵母细胞 绵羊 |
| 收稿时间： | 2009/3/9 0:00:00 |
| 修稿时间： | 2009/4/1 0:00:00 |
Influential Factors of in Vitro Maturated Ovine Oocytes Cryopreserved by GMP on the Developmental Competence |
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| QUAN Fusheng,LIU Yingxun,LIU Lijie,SHEN Yu,MA Huiming,YU Yuan and ZHANG Yong.Influential Factors of in Vitro Maturated Ovine Oocytes Cryopreserved by GMP on the Developmental Competence[J].Acta Agriculturae Boreali-occidentalis Sinica,2009,18(6):18-22. |
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| Authors: | QUAN Fusheng LIU Yingxun LIU Lijie SHEN Yu MA Huiming YU Yuan and ZHANG Yong |
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| Institution: | The Medicine and Veterinary College, Northwest A&F University, Yangling Shaanxi 712100, China;The Medicine and Veterinary College, Northwest A&F University, Yangling Shaanxi 712100, China;The Medicine and Veterinary College, Northwest A&F University, Yangling Shaanxi 712100, China;The Medicine and Veterinary College, Northwest A&F University, Yangling Shaanxi 712100, China;The Medicine and Veterinary College, Northwest A&F University, Yangling Shaanxi 712100, China;The Medicine and Veterinary College, Northwest A&F University, Yangling Shaanxi 712100, China;The Medicine and Veterinary College, Northwest A&F University, Yangling Shaanxi 712100, China |
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| Abstract: | In the present study, in vitro maturated ovine oocytes were cryopreserved by GMP (glass micropipette vitrification), and the developmental capacity of ovine oocytes cryopreserved by different frozen-thawed protocols was investigated. Effects of addition of 0. 3mol/L sucrose in cryoprotectants, pre-treatment with CB before vitrification, removing cumulus cells of oocytes and stepwise vitrified-warmed oocytes with three steps or four steps manner on the morphological normal rate and developmental rate of parthenogenetic embryos were observed. The cleavage rates (56. 70%) of the partheno-genetic embryos of the addition of sucrose group were significantly higher than that of control group (25.40%) (P<0. 01), and the blastocyst rates (16.49%) were higher than that of control (4.76%) (P<0. 05); the morphological normal rates of the CB pretreatment group were higher than that of control groups (61. 11%) (P<0. 05), the cleavage rates (39. 39%) were also higher than that control groups (21. 11%); there were not significant differences in the morphological normal rates and blastocyst rates between nude oocyte and COCs to be cryopreserved; the groups of the four-step were not different from the groups of the three-step in the morphological normal rates and the cleavage rates of parthenogenetic embryos when thawed (P<0. 05). The addition of sucrose in the vitrification solution and pretreatment with CB before vitrification had advantage of the morphological returned to normal and development capacity of the oocytes cryopreserved by GMP. |
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| Keywords: | GMP |
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