在玻璃化培养基中添加维生素C和氯化锌对玻璃化未成熟牛卵母细胞活力、体外成熟和超微结构变化的影响

H. Yassen, K. Attia, S. Shamiah, M. El-Arian, M. El-Harairy
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摘要

本研究旨在探讨在玻璃化培养基中添加维生素C和氯化锌对玻璃化巴拉地牛未成熟卵母细胞体外成熟活力和超微结构变化的影响。从屠宰牛卵巢中获得致密积云卵母细胞(COCs) (n=1370)。用体视显微镜观察卵母细胞形态。台盼染色观察体外成熟及超微结构变化。结果表明,氯化锌玻璃化的牛卵母细胞总存活率为90.28%,正常存活率为81.11%,显著高于维生素C玻璃化的82.5%,正常存活率为65.65%,高于对照的74.44%,正常存活率为54.72%。异常牛卵母细胞的恢复率(9.17 vs. 16.94和19.72%)呈显著相反趋势。与添加维生素C和未添加维生素C的培养基(74.55%和68.63%)相比,新鲜培养基(对照)中活细胞质和活卵丘的卵母细胞比例(93.75%,P<0.05)显著提高(P<0.05)。在可变细胞质和可变积云(VOUC)的卵母细胞中,氯化锌、对照和维生素C培养基之间差异不显著。添加氯化锌和维生素C的玻璃化培养基比不添加氯化锌和维生素C的玻璃化培养基显著(P<0.05)提高了卵丘复合体(COCs)的成熟率(MII)。在不添加维生素C、不添加氯化锌的情况下,玻璃化处理的牛卵母细胞大部分细胞器超微结构改变率显著升高(P<0.05)。结论:在玻璃化培养基中添加维生素C或氯化锌可提高牛卵母细胞的存活率、形态和超微结构以及成熟率
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Effect of Addition Vitamin C and Zinc Chloride in Vitrification Medium on Viability, in vitro Maturation and Ultrastructure Changes of Vitrified Immature Bovine Oocytes
This study aimed to evaluate the impact of adding vitamin C and zinc chloride to vitrification medium on viability in vitro maturation and ultrastructure changes of vitrified immature Baladi cow oocytes. Compact cumulus oocytes (COCs) (n=1370) were obtained from slaughtered bovine ovaries. Then the morphology of oocytes was examined using a stereomicroscope. Staining with trypan, the in vitro maturation and ultrastructural changes were studied. The results revealed significant (P<0.05) increase of total and normal survival rate of bovine oocytes vitrified with zinc chloride (90.28 and 81.11%) than in vitamin C media (82.5 and 65.65%) or control medium (74.44 and 54.72%). Recovery rate of abnormal bovine oocytes showed significantly an opposite trend (9.17 vs. 16.94 and 19.72%). Proportion of oocytes with viable cytoplasm and viable cumulus (VOVC) was increased significantly (93.75%, P<0.05) in fresh (control) than in medium supplemented with vitamin C and untreated medium (74.55 and 68.63%), respectively. There were non-significant differences among zinc chloride, control and vitamin C media in oocytes with vaible cytoplasm and unvaiable cumulus (VOUC). Supplementation of the vitrification medium with zinc chloride and vitamin C significantly (P<0.05) improved maturation rates (MII) of recovered cumulus oocyte complexes (COCs) than medium without supplementation. The percentage of ultrastructural alterations in most organelles bovine oocytes significantly (P<0.05) increased in oocytes vitrified without supplementation followed by vitamin C, then zinc chloride medium. Conclusion, supplementation of vitamin C or zinc chloride to the vitrification medium improved survival rate, morphologically and ultrastructural, as well as maturation rate of bovine oocytes
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