L. Dujíčková, L. Olexiková, E. Kubovicova, J. Bezdíček, M. Ješeta, A. Makarevich
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引用次数: 0
摘要
摘要本研究的目的是检测冷冻保存后牛卵母细胞的活力。体外成熟(IVM)后的卵母细胞通过超快速冷却技术在镍电子显微镜网格上以最小体积玻璃化。在加温和随后的体外受精后,将推定受精卵培养至胚泡阶段(Bl)。对几个失透的卵母细胞进行了电镜分析。尽管玻璃化组的胚胎切割和Bl百分比略低于对照组(P<0.05),但两组的Bl总细胞数(TCN)、凋亡和死细胞百分比没有差异。然而,在对照组(108.0 vs 90.5)和玻璃化组(103.75 vs 98.14)的TCN第7天(D7)和第8天(D8)Bl之间发现了显著差异。然而,冷冻卵母细胞的电子显微镜显示线粒体和滑面内质网(SER)有轻微的可逆损伤,失透卵母细胞发育到Bl期与新鲜卵母细胞相当。总之,与D7-Bl相比,发育较慢的Bl(D8)比例更高,这可能与玻璃化卵母细胞中某些细胞器的轻微损伤有关。
Viability of bovine in vitro matured oocytes following ultra-rapid vitrification
Abstract The aim of the study was to examine viability of cattle oocytes after cryopreservation. Oocytes after in vitro maturation (IVM) were vitrified in minimum volume on the nickel electron microscopy grids by ultra-rapid cooling technique. After warming and subsequent in vitro fertilization the presumptive zygotes were cultured to reach the stage of the blastocyst (Bl). Several devitrified oocytes were processed for electron microscopy assay. Although, embryo cleavage and Bl percentages in the vitrified group were slightly lower than in the control group (P < 0.05), the Bl total cell number (TCN), apoptosis and dead cell percentages did not differ between both groups. However, significant difference was found between day 7 (D7) and day 8 (D8) Bl in the TCN in control (108.0 vs. 90.5) and vitrified group (103.75 vs 98.14). Electron microscopy of frozen oocytes revealed slight reversible injuries in mitochondria and the smooth endoplasmic reticulum (SER), nevertheless, the development of devitrified oocytes to the Bl stage was comparable to those in fresh oocytes. In conclusion, higher proportion of slower developing Bl (D8) compared to D7 Bl may be related to the mentioned minor damages of some organelles in vitrified oocytes.