【作者】 庞春英； 杨春艳； 李日聪； 黄健； 黄芬香； 郑海英； 覃广胜； 张秀芳； 杨炳壮； 梁贤威；

【Author】 Chunying Pang;Chunyan Yang;Richong Li;Jian Huang;Fenxiang Huang;Haiying Zheng; Guangsheng Qin;Xiufang Zhang;Bingzhuang Yang;Xianwei Liang~★Guangxi Buffalo Research Institute,Chinese Academy of Agricultural Sciences,Nanning city,Guangxi 530001

【机构】 中国农业科学院广西水牛研究所；

【摘要】 本研究采用不同来源的体外生产胚胎(屠宰场卵巢IVF胚胎,OPU-IVF胚胎,SCNT胚胎),系统研究探讨了不同冷冻方法对水牛体外生产胚胎冷冻效果的影响,以完善水牛体外生产胚胎冷冻方法,进一步提高胚胎冷冻效果。试验选用6-7日龄囊胚分别用不同的冷冻液和不同冷冻方法进行胚胎冷冻。玻璃化冷冻液分别为40%EG、25%EG+25%DMSO和20%EG+20%DMSO+0.5 M蔗糖;程序化冷冻液分别为10%甘油和0.05M海藻糖+1.8M EG+0.4%BSA。结果表明:(1)在玻璃化冷冻中,无论何种胚胎不同冷冻液的冷冻效果有明显的差异,但均以20%EG+20%DMSO+0.5 M蔗糖作为冷冻液的冷冻效果最好,并且高于程序化冷冻的存活率;而对于程序化冷冻,用1 0%甘油作为冷冻液,屠宰场卵巢IVF胚胎的冷冻后存活率略高于用0.05M海藻糖+1.8M EG+0.4%BSA的冷冻后存活率,但差异不显著(P>0.05)。(2)程序化冷冻胚胎解冻后,在0-24 h内有76.5%胚胎复活,高于玻璃化冷冻的复苏率(48.9%)(p<0.05);而与此相反,在24-48 h内,玻璃化冷冻胚胎的复苏率(42.6%)则高于程序化冷冻(2 3.5%)(p<0.05)。综上所述,应用玻璃化冷冻的效果好于程序化冷冻,且以20%EG+20%DMSO+0.5 M蔗糖作为冷冻液进行玻璃化冷冻效果最好,但程序化冷冻后的胚胎复苏速度明显快于玻璃化冷冻的速度。更多还原

【Abstract】 The Days 6-7 buffalo SCNT and the IVF blastocysts derived from abattoir ovaries and OPU were cryopreserved by slow freezing and vitrification in different cryoprotectant mediums.40%EG,25%EG+ 25%DMSO and 20%EG+20%DMSO+0.5 M sucrose were adopted for vitrification,while 10%glycerol and 0.05M trehalose dehydrate+1.8%EG+0.4%BSA were used for slow freezing.The results showed that(1):the efficiency of different vitrification medium was different from each other,20%EG+20%DMSO+0.5 M sucrose showed the best result which was also higher than the slow freezing mediums.As for slow freezing,the efficiency of the two kinds of medium were not different from each other.(2):76.5%of the slow frozen blastocysts survived in 0-24 h after thawing,which was higher than that of vitrified blastocysts(48.9%).On the contrary,more vitrified blastocysts(42.6%) survived in 24-48 h than the slow frozen blastocysts(23.5%).In conclusion,the efficiency of vittification is higher than slow freezing,and the medium of 20%EG+20%DMSO+0.5 M sucrose is best,while the slow frozen blastocysts survived earlier than the vitrified ones after thawing.更多还原