Survival and apoptosis rates after vitrification in cryotop devices of in vitro-produced calf and cow blastocysts at different developmental stages
Survival and apoptosis rates after vitrification in cryotop devices of in vitro-produced calf and cow blastocysts at different developmental stages
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DOI：
10.1071/rd10013
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发表时间：
2010-01-01
影响因子：
1.9
通讯作者：
Mogas, Teresa
中科院分区：
文献类型：
--
作者：
Morato, Roser;Izquierdo, Dolors;Mogas, Teresa
设计了两个实验来确定处于不同发育阶段的体外培养的囊胚在使用cryotop装置的玻璃化冷冻程序中存活的能力。第7天和第8天的胚胎被分类为非扩张、扩张或孵化和/或孵化的囊胚。在第一个实验中，我们研究了玻璃化温热囊胚在合成输卵管液（SOF）培养基中孵育3小时后的存活率。在第二个实验中，使用末端脱氧核糖核苷酸转移酶介导的dUTP-洋地黄毒苷缺口末端标记（TUNEL）技术检测DNA受损的细胞核，对玻璃化温热的囊胚进行评价。在这两个实验中，比较了奶牛和小牛囊胚的结果。玻璃化冷冻第8天扩张（52.4%）和孵化（50%）的奶牛囊胚或第8天扩张（54.5%）和孵化（59.4%）的小牛囊胚后，未观察到存活率差异。当胚胎在第7天玻璃化冷冻时，观察到在加温扩张和孵化后的牛囊胚的存活率分别为78.4%和66.7%，而小牛囊胚的存活率分别为80%和76.9%。与其他发育阶段相比，非扩张囊胚的存活率最低（26%-54%），特别是第8天玻璃化冷冻的囊胚（
Two experiments were designed to determine the ability of in vitro-cultured blastocysts at different stages of development to survive the vitrification procedure using cryotop devices. Day 7 and Day 8 embryos were classified as non-expanded, expanded or hatching and/or hatched blastocysts. In the first experiment, we examined the survival rate of vitrified-warmed blastocysts after 3 h incubation in synthetic oviducal fluid (SOF) medium. In the second experiment, vitrified-warmed blastocysts were evaluated using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) technique to detect nuclei with damaged DNA. In both experiments, results for cow and calf blastocysts were compared. No differences in survival rates were observed after vitrification of Day 8 expanded (52.4%) and hatched (50%) cow blastocysts or Day 8 expanded (54.5%) and hatched (59.4%) calf blastocysts. When embryos were vitrified on Day 7, survival rates of 78.4% and 66.7% were observed after warming expanded and hatched cow blastocysts, respectively, compared with rates of 80% and 76.9%, respectively, for calf blastocysts. Lowest survival rates were recorded for non-expanded blastocysts (26%-54%) compared with the other developmental stages, particularly those vitrified at Day 8 (