Cryopreservation of mature bovine oocytes by vitrification in straws
Cryopreservation of mature bovine oocytes by vitrification in straws
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DOI：
10.1006/cryo.1998.2103
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发表时间：
1998-08-01
期刊：
影响因子：
2.7
通讯作者：
Suzuki, T
中科院分区：
文献类型：
--
作者：
Otoi, T;Yamamoto, K;Suzuki, T
进行实验以确定体外成熟牛卵母细胞在细管中玻璃化冷冻的最佳条件。在第一个系列的实验中，测试了在将卵母细胞暴露于由 30% (v/v) 乙二醇 (EG) 和 0.35 M 蔗糖组成的玻璃化溶液中之前逐步添加的效果。三步添加法玻璃化冷冻卵的形态存活率和卵裂率均高于一步添加法玻璃化冷冻卵母细胞。在第二个系列中，测试了玻璃化溶液浓度的影响（20%、30%、40% 和 50% EG，均含有 0.35 M 蔗糖）。 20%和30%EG玻璃化冷冻卵母细胞的存活率低于40%和50%EG玻璃化冷冻的卵母细胞存活率。 40% EG 玻璃化冷冻卵母细胞的卵裂率和囊胚发育率是四组中最高的。在第三个系列中，测试了三步稀释过程中第一步卵母细胞暴露于 0.5 M 蔗糖的持续时间（1 分钟、5 分钟和 10 分钟）的影响。虽然在 40% EG 中玻璃化冷冻的卵母细胞的发育能力方面，各组之间没有显着差异，但当卵母细胞暴露于蔗糖 5 分钟时，观察到最高的囊胚发育率 (10%)。这些数据表明，冷冻保护剂浓度和添加程序的改进对玻璃化细管中卵母细胞的发育能力具有关键影响。 (C) 1998 年学术出版社。
Experiments were conducted to determine optimal conditions for vitrification of in vitro matured bovine oocytes in straws. In the first series of experiments, the effects of stepwise addition before exposure of oocytes to vitrification solution consisting of 30% (v/v) ethylene glycol (EG) with 0.35 M sucrose was tested. The rates of morphological survival and cleavage of oocytes vitrified by the three-step addition procedure were higher than those vitrified by the one-step addition procedure. In the second series, the effect of the concentration of the vitrification solution was tested (20, 30, 40, and 50% EG, all with 0.35 M sucrose). The survival rates of oocytes vitrified in 20 and 30% EG were lower than those in 40 and 50% EG. The rates of cleavage and development to blastocysts of oocytes vitrified in 40% EG were the highest among the four groups. In the third series, the effect of duration of exposure of oocytes to 0.5 M sucrose (1, 5, and 10 min) at the first step during the three-step dilution was tested. Although there were no significant differences among the groups with respect to developmental competence of oocytes vitrified in 40% EG, the highest development rate (10%) to blastocysts was observed when oocytes were exposed to sucrose for 5 min. These data demonstrate that improvements to the concentration of cryoprotectant and addition procedures have critical effects on the developmental competence of oocytes vitrified in straws. (C) 1998 Academic Press.