不同浓度玻璃化冷冻液对牛未成熟卵母细胞发育能力的影响

摘要

The aim of this study was to evaluate the effect of different vitrification solutions on the potential development competent of immature oocytes in bovine.In experiment 1,cumulus oocyte complexes (COCs) were placed in vitrification solution 1 (VS1,10％ EG+ 10％ DMSO) or vitrification solution 2 (VS2,20％ EG+20％ DMSO) or vitrification solution 3(VS3,25 EG+25％ DMSO) for either 30 or 60 s.There were no differences among VS1,VS2 and control group for maturation rates(P＞0.05).However,maturation rate of 60 s in VS3 was decreased than that of the control group(P＜0.01).In experiment 2,COCs were cryopreseved in open pulled straw (OPS) in different solutions of VS1 or VS2 or VS3.The highest blastocyst rate (5.4％) was in VS2,the lowest was in VS1 (0),there were no defferences between VS2 and VS3 (3.6％) groups.It can be concluded that solution contained 20％ EG and 20％ DMSO can be used for vitrification of immature bovine oocytes,and the exposing time of COCs in the cryoprotectant should be limited in 30 s.%旨在研究含不同浓度乙二醇(EG)、二甲基亚砜(DMSO)的玻璃化冷冻液对牛未成熟卵母细胞冷冻后发育能力的影响.将卵丘卵母细胞复合体(COCs)置于不同配方的玻璃化冷冻液(VS1:10％EG+10％DMSO; VS2:20％EG+20％DMSO; VS3:25％EG+25％DMSO)中暴露30 s或60 s,体外成熟培养22 h,以未在玻璃化冷冻液中暴露而直接体外成熟22 h的COCs为对照组.结果表明,在VS3中暴露60 s时,成熟率比对照组明显下降(P＜0.01),其他组与对照组差异均不显著(P＞0.05);在不同玻璃化冷冻液(VS1、VS2、VS3)中,采用开放式拉长细管(OPS)法冷冻COCs,解冻后再进行体外成熟(IVM)、体外受精(IVF)和早期胚胎的体外培养(IVC).结果表明,VS2组囊胚率(5.4％)最高,与VS3组(3.6％)差异不显著(P＞0.05),VS1组没有获得囊胚.研究表明,含20％ EG、20％ DMSO的玻璃化冷冻液可以用于牛未成熟卵母细胞的冷冻,COCs在冷冻液中处理的时间应控制在30s内.