玻璃化冷冻保存对体外成熟猪卵母细胞DNA的影响  

Porcine Oocytes Vitrification-Induced DNA Damage Measured with Single Cell Gel Electrophoresis

作　　者：肖宇[1] 蒋伟娟 唐峰[1] 朱淑文[1] 徐动[1] 

机构地区：[1]上海交通大学农业与生物学院,上海200240 [2]上海市兽医卫生监督所,上海200335

出　　处：《上海交通大学学报（农业科学版）》2008年第1期50-54,共5页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：上海市科委科学发展基金(No:064909004)

摘　　要：采用乙二醇(EG)+二甲基亚砜(DMSO)作为冷冻保护剂处理并使用微管法冷冻保存猪卵母细胞,运用细胞形态观察和单细胞凝胶电泳(SCGE)方法检测玻璃化冷冻过程对细胞DNA损伤情况。结果显示:保护剂处理冷冻组,细胞形态正常率和DNA异常率分别为63.364%、42.059%;同保护剂处理未冷冻对照组82.571%、25.758%的差异显著(P<0.05)。表明冷冻保存过程对猪卵母细胞DNA有一定影响;以无冷冻保护剂处理卵母细胞为对照检测保护剂的细胞毒性,对照组形态正常率和DNA异常率分别为84.51%、12.28%,与冷冻保护剂处理组有显著差异(P<0.05)。说明冷冻保护剂对猪卵母细胞DNA有明显细胞毒性作用。The effects of vitrification-induced DNA damage and toxicity of vitrificational solution which contains ethylene glycol （EG） and dimethylsulfoxide（DMSO） on in vitro maturation （IVM） porcine oocytes were detected by single cell gel electrophoresis （SCGE）.The vitrificared thawed oocytes and the control unfrozen oocytes were immediately subjected to morphological evaluation and underwent comet assay （SCGE） to detect cryoinjury at DNA level which caused by vtrrification in 0.25 mL straws.The results showed the morphology intact rates and DNA damage rates of vitrificated oocytes were 63.364% and 42.059%, and the control which unfrosen were 82.571%,25.758 %,there is significant morphological damage as well as DNA damage compared to unfrozen control （P〈0.05）. Oocytes that had been treated with vitrificational solution exhibited more DNA damage than the control without treated which morphology intact rates and DNA damage rates were 84.510% and 12.28%.This experiment demonstrated that oocyte DNA is a target of cryoinjury and the vitrificational solution result in toxicity for porcine oocytes to some extent.

关 键 词：猪卵母细胞 玻璃化冷冻保存 单细胞凝胶电泳 DNA损伤 CASP分析 

分 类 号：Q954.1[生物学—动物学]