Chasombat Jakkhaphan, Vongpralub Thevin, Sirisathien Saksiri, Phasuk Yupin, Sonseeda Pronjit
Cryo Letters. 2015 May-Jun;36(3):165-73.
The present study aimed to improve the oocyte vitrification procedure for preservation of Thai native cattle genetic resources. In Experiment I, oocytes were exposed to various doses (2%, 4% and 6%) of ethylene glycol (EG) in vitrification solution I (VS-I) for different equilibration times (10 or 20 min) before being exposed to VS-II and then subjected to vitrification. Experiment II was divided into two parts: (a) oocytes were matured in medium supplemented with linoleic acid albumin (LAA) (1% or 2%) and then vitrified; (b) matured oocytes were preincubated with cholesterol-loaded methyl-β-cyclodextrin (CLC) (1% or 2%) and then vitrified. Equilibration of oocytes by exposure to 6% EG in VS-I for 10 min (Experiment I), and in vitro maturation of immature oocytes in medium supplementation with 2% LAA (Experiment II) were the most effective methods; vitrified/thawed oocytes showed higher rates of survival and subsequent embryonic development compared with the other experimental groups.
本研究旨在改进用于保存泰国本土牛遗传资源的卵母细胞玻璃化程序。在实验I中，卵母细胞在玻璃化溶液I（VS-I）中暴露于不同剂量（2%、4%和6%）的乙二醇（EG），平衡不同时间（10或20分钟），然后再暴露于VS-II并进行玻璃化处理。实验II分为两部分：（a）卵母细胞在添加亚油酸白蛋白（LAA）（1%或2%）的培养基中成熟，然后进行玻璃化处理；（b）成熟卵母细胞先用载胆固醇的甲基-β-环糊精（CLC）（1%或2%）预孵育，然后进行玻璃化处理。在VS-I中用6% EG处理10分钟对卵母细胞进行平衡（实验I），以及在添加2% LAA的培养基中对未成熟卵母细胞进行体外成熟（实验II）是最有效的方法；与其他实验组相比，玻璃化/解冻后的卵母细胞显示出更高的存活率和随后的胚胎发育率。
