GMP玻璃化冷冻对水牛MⅡ期卵母细胞冷冻损伤和体外发育能力的影响  

Effects of GMP Vitrification on the Cold Injure and in vitro Fertilization of MetaphaseⅡ Buffalo Oocytes

作　　者：杨素芳[1] 陈华海[1] 蒋建荣[1] 冯贵学[2] 陈焕华[1] 陈玉[1] 卞桂华[1] 石德顺[1] 

机构地区：[1]广西大学动物繁殖研究所,广西南宁530005 [2]广西壮族自治区妇幼保健院生殖中心,广西南宁530003

出　　处：《河南农业科学》2009年第1期104-108,共5页Journal of Henan Agricultural Sciences

基　　金：国家“863”计划项目(2007AA100505);广西青年基金项目(桂科青0542022)

摘　　要：将体外成熟(IVM)24h的水牛卵母细胞随机分成对照组、冷冻剂毒性试验组、GMP玻璃化冷冻组,研究玻璃微细管法(GMP)冷冻对水牛MⅡ期卵母细胞冷冻损伤和体外受精后发育能力的影响。结果表明:冷冻剂毒性试验组的存活率显著高于GMP玻璃化冷冻组;在透明带消化时间上,冷冻剂毒性试验组和GMP玻璃化冷冻组显著高于对照组。在单精入卵率上,冷冻剂毒性试验组和GMP玻璃化冷冻组显著低于对照组;在无精子入卵率上,冷冻剂毒性试验组和GMP玻璃化冷冻组与对照组差异显著;而多精入卵率3组间差异不显著。卵母细胞冷冻后进行体外受精,3组之间的2-细胞率,8-细胞率和桑椹胚率差异均极显著。表明冷冻剂和GMP玻璃化冷冻均会引起水牛MⅡ期卵母细胞透明带硬化,降低精子穿透率和其体外受精的发育能力。To investigate the effects of glass microscopic grids （GMP） vitrification on the cold in- jure and in vitro fertilization of MetaphaseⅡ （M Ⅱ ） buffalo oocytes, the oocytes were randomly allocated into three groups after in vitro maturation for 24h： （1） control group, （2） toxicity group exposed to vitrification solution without being plunged into liquid nitrogen, （3） vitrification group vitrified by GMP method. The results showed that the survival rate of toxicity group was significantly higher than of vitrification group （P^0.01）. The zona pelludica digestion duration in the oocytes of the toxicity group and vitrification group was longer than that of control group （P〈0. 05）. The percentage of monospermy in the toxicity group and vitrification group was significantly decreased compared with that of the control group （P〈0. 05）. The percentage of non-penetration in the toxicity group and vitrification group was higher than that of control group （P〈0.05）. The percentage of polyspermy had no significantly difference around the three groups （P〉0.05）. The difference among 2-cell rate, 8-cell rate and the morula rate of the control group, toxicity group and vitrification group were extremely significant （P〈0. 01）. These data indicated that both of the vitrification solution and vitrification by GMP resulted in zona peltudica harding of M Ⅱ buffalo oocytes, which reduced the sperm penetration and decreased the development ability of oocytes after fertilizated in vitro.

关 键 词：水牛 MⅡ期卵母细胞 GMP 体外受精 

分 类 号：S814.8[农业科学—畜牧学]