| 不同冷冻方法对水牛体外生产胚胎冷冻效果的研究 |
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| 引用本文： | 庞春英,杨春艳,杨炳壮,李日聪,黄健,黄芬香,郑海英,覃广胜,张秀芳,梁贤威.不同冷冻方法对水牛体外生产胚胎冷冻效果的研究[J].畜牧与兽医,2011,43(5). |
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| 作者姓名： | 庞春英 杨春艳 杨炳壮 李日聪 黄健 黄芬香 郑海英 覃广胜 张秀芳 梁贤威 |
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| 作者单位： | 中国农业科学院广西水牛研究所广西水牛遗传繁育重点实验室,广西南宁,530001 |
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| 基金项目： | 农业部948项目，科技部国际合作项目2008DFA30320，广西科技攻关项目 |
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| 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 mol/L蔗糖;程序化冷冻液分别为10%甘油和0.05 mol/L海藻糖+1.8 mol/L 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 mol/L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10%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0.05 mol/L海藻糖+1.8 mol/L EG+0.4%BSA的冷冻后存活率,但差异不显著(P>0.05)。(2)VF胚胎利用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(23.5%)(P<0.05)。综上所述,各种来源的水牛体外生产胚胎均可进行冷冻保存,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 mol/L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。
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| 关 键 词： | 水牛 体外生产 胚胎 玻璃化冷冻 程序化冷冻 |
Effects of different methods on cryopreservation of embryos produced in vitro in buffalo |
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| PANG Chun-ying,YANG Chun-yan,YANG Bing-zhuang,LI Ri-cong,HUANG Jian,HUANG Fen-xiang,ZHENG Hai-ying,QIN Guang-sheng,ZHANG Xiu-fang,LIANG Xian-wei.Effects of different methods on cryopreservation of embryos produced in vitro in buffalo[J].Animal Husbandry & Veterinary Medicine,2011,43(5). |
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| Authors: | PANG Chun-ying YANG Chun-yan YANG Bing-zhuang LI Ri-cong HUANG Jian HUANG Fen-xiang ZHENG Hai-ying QIN Guang-sheng ZHANG Xiu-fang LIANG Xian-wei |
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| Institution: | PANG Chun-ying,YANG Chun-yan,YANG Bing-zhuang,LI Ri-cong,HUANG Jian,HUANG Fen-xiang1,2,ZHENG Hai-ying,QIN Guang-sheng,ZHANG Xiu-fang,LIANG Xian-wei(Guangxi Buffalo Research Institute,Chinese Academy of Agricultural Science,Guangxi Key Laboratory of Buffalo Genetics,Breeding and Reproduction,Nanning 530001,China) |
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| Abstract: | 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.The 40%EG,25%EG+25%DMSO and 20%EG+20%DMSO+0.5 mol/L sucrose were adopted for vitrification,while 10% glycerol and 0.05 M 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 mol/L sucrose was the best.A... |
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| Keywords: | buffalo in vitro production embryo vitrification slow freezing |
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