摘要
本实验旨在探讨玻璃化冷冻保存对猪MⅡ期卵母细胞皮质颗粒分布和孤雌激活后早期胚胎发育能力的影响。实验将卵母细胞随机分为对照组、毒性实验组和冷冻组。采用EFS40和EDFS40两种玻璃化冷冻液处理,卵母细胞经恢复后对其进行染色,观察皮质颗粒的分布;并对另一部分卵母细胞实施孤雌激活,观察早期胚胎的发育。结果表明:毒性实验组和冷冻组卵母细胞皮质颗粒部分释放、完全释放的比例无显著差异,但均显著高于对照组(P<0.05)。不同毒性处理组和不同冷冻组间对皮质颗粒的分布无显著差异。与毒性实验组相比,冷冻处理组显著降低皮质颗粒在皮质区分布的比例(P<0.05)。EFS40毒性实验组孤雌激活后的存活率、卵裂率、囊胚发育率均显著高于EDFS40毒性实验组(86.6%vs.75.0%)、(61.8%vs.40.7%)、(30.2%vs.23.5%)(P<0.05)。EFS40冷冻组存活率显著高于EDFS40冷冻组,但均显著低于对照组。结果显示,抗冻保护剂处理和玻璃化冷冻均导致猪卵母细胞皮质颗粒释放,与EDFS40相比采用EFS40较适合猪MⅡ期卵母细胞冷冻保存。
This study was designed to investiga te the effects of vitrification on cortical granulesdistribution of in vitro matured porcine oocytes and evaluate their embryonic developmental competence after parthenogenetic activation.Two kinds of vitrification solution EDF40 and EDFS40 were used.Oocytes were randomly divided into three groups： control group,toxicity groups,vitrification groups.Part of treated oocytes were stained to observe the distribution of cortical granules.The rest were cultured in vitro to assess their developmental competence after parthenogenetic activation.The results showed that there were no significant differences in proportion of cortical granules partially release distribution and fully release distribution between toxicity groups and vitrification groups,but their proportions were significantly higher than that in control group（P 〈0.05）.There were no significant differences between different toxicity groups or vitrification groups.The proportions of cortical granule distributed in the cortex in vitrification groups were significantly lower than that in toxicity groups（P〈0.05）.The viability rates,cleavage rages and blastocyst rates in EFS40 toxicity group were significantly higher than EDFS40 toxicity group（86.6% vs.75.0%）,（61.8% vs.40.7%）,（30.2% vs.23.5%）（P 〈0.05）.The viability rates of EFS40 vitrification group was significantly higher than those of EDFS40 vitrification group,but they were both significantly lower than that in the control group.In conclusion,cryoprotectants and vitrification lead to the release of cortical granules of porcine oocytes,EFS40 is more suitable than EDFS40 for cryopreservation in vitro matured porcine oocytes.
出处
《中国畜牧杂志》
CAS
北大核心
2012年第19期29-32,共4页
Chinese Journal of Animal Science
基金
国家863项目(2011AA100303)
国家科技支撑项目(2011BAD19B01)
