不同温度对犬卵母细胞体外卵丘扩散及核成熟的影响  

Effect of Incubation Temperature on in vitro Maturation of Canine Oocytes:Cumulus Cell Expansion and Nuclear Maturation

作　　者：李所[1] 康锦丹[1] 金君学[1] 洪禹[1] 朱海瑛[1] 金龙[1] 郭庆[1] 尹熙俊[1] 

机构地区：[1]延边大学农学院动物科学系,吉林延吉133002

出　　处：《中国畜牧杂志》2014年第3期35-38,共4页Chinese Journal of Animal Science

基　　金：973计划前期研究专项(2012CB526710);转基因动物与胚胎工程吉林省重点实验室平台建设

摘　　要：目前犬体外胚胎生产及克隆等技术由于受到卵母细胞体外成熟率低的影响而受到限制,实验旨在探索不同温度对犬卵母细胞体外卵丘扩散及核成熟的影响。从不同时期犬卵巢收集卵丘卵母细胞复合体(COCs),随机分成4组,置于TCM-199(Earle’s salts)+25 mmol/L HEPES+10%(v/v)FBS+10 ng/mL EGF+1 mg/mL L-半胱氨酸+1%青–链霉素成熟培养液中,分别在37.0、38.5、40.0℃条件下培养48 h,测定不同温度培养前后卵丘扩散及核成熟情况。结果表明:不同温度下,培养前后卵丘均显著性扩散(211.40μmvs359.64、377.25、373.23μm),但各处理组间无显著差异,不同温度下对犬卵母细胞核体外成熟率(10.73%、10.025%、14.35%)没有显著差异。说明培养温度只要在犬正常体温变动范围内,则温度对其卵母细胞卵丘扩散及体外培养成熟率均无显著影响。At present, canine embryos production in vitro and cloning technology were limited by the low in vitro maturation rate of canine oocyte. The objective of this study was to evaluate the effect of incubation temperature on in vitro maturation of canine oocytes： cumulus cell expansion and nuclear maturation. Cumulus oocyte complexes （COCs） were harvested from ovaries and matured in TCM199 medium buffered with 25 mmol/L HEPES buffer supplemented with 10% （v/v）FBS, 10 ng/mL epidermal growth factor （EGF）, 1 mg/mL cysteine and 1% penicillin-streptomycin. Oocytes were cultured at 37.0℃, 38.5℃, 40.0℃ in a humidified atmosphere of 5% CO2 for 48 h incubator, respectively. After culture, oocyte were stained and examined for nuclear maturation. The results showed that there were no differences among treatments in the cumulus cell expansion （211.40 μm vs 359.64 μm, 377.25 μm, 373.23 μm） and nuclear maturation （10.73%, 10.02%, 14.35%）. As long as the incubation temperature within the range of dogs normal body temperature, there were no significant differences among treatments.

关 键 词：卵母细胞 温度 体外成熟 犬 

分 类 号：S829.202[农业科学—畜牧学]