温度和冻存液体积与冷冻液接触时间对双层麦管法冻存小鼠卵子的影响  

Effects of temperature,volume of frozen liquid and contact time of frozen liquid on frozen mouse eggs by double-tube method

作　　者：张靖 陶娟 张晗 杨杰 高璐 韩梦雅 ZHANG Jing;Tao Juan;ZHANG Han;YANG Jie;GAO Lu;HAN Mengya(Sanquan College of Xinxiang Medical University,Xinxiang,Henan,453003,China)

机构地区：[1]新乡医学院三全学院,河南新乡453003

出　　处：《当代医学》2023年第11期1-4,共4页Contemporary Medicine

基　　金：河南省科技攻关项目(202102310386);河南省高等学校重点科研项目(20B180011);河南省高校青年骨干教师资助计划(2017GGJS220)。

摘　　要：目的探讨温度和冻存液体积与冷冻液接触时间对双层麦管法冻存小鼠卵子的影响。方法90只KM品系雌鼠注射激素,使其超数排卵,以获得形态良好的带放射冠的卵子,采用双麦管法玻璃化冷冻与复苏带放射冠的卵子。根据不同操作温度(22℃、37℃)、冷冻液接触时间(>30~40、>40~90、90~180 s)、冻存液体积(>0.1~0.5μl、>0.5~1.0μl)分别冷冻,比较各种冷冻条件下的卵子存活率。结果操作时间为40~90 s,冻存液体积为0.1~0.5μl时,22℃卵子存活率高于37℃,差异有统计学意义(P<0.05)。22℃环境下,冻存液体积为>0.1~0.5μl时,>40~90 s操作时间小鼠卵子存活率高于>30~40、>90~180 s,>90~180 s操作时间小鼠卵子存活率高于>30~40 s,差异有统计学意义(P<0.05)。在22℃环境下,操作时间为>40~90 s时,>0.1~0.5μl冻存液体积小鼠卵子存活率高于>0.5~1.0μl,差异有统计学意义(P<0.05)。结论应选择22℃环境温度,>40~90 s的操作时间,>0.1~0.5μl的冻存液体积冻存带有放射冠的小鼠卵子,效果最佳,存活率最高。Objective To explore the effects of temperature,volume of frozen liquid and contact time of frozen liquid on frozen mouse eggs by double-tube method.Methods 90 KM line female mice were injected with hormones to make them superovulate and obtain well-formed ovum with radiation crown.According to different operating temperature(22℃,37℃),refrigerant contact time(>30-40,>40-90,>90-180 s),frozen liquid volume(>0.1-0.5μl,>0.5-1.0μl),respectively.The survival rate of eggs under different freezing conditions was compared.Results When the operation time was 40-90 s and the frozen fluid volume was 0.1-0.5μl,the egg survival rate at 22℃was higher than that at 37℃,and the difference was statistically significant(P<0.05).At 22℃,when the frozen liquid volume was>0.1-0.5μl,the egg survival rate of mice during>40-90 s was higher than that of>30-40 and>90-180 s,and the egg survival rate of mice during>90-180 s was higher than that of 30 to 40 s,the difference was statistically significant(P<0.05).When the operation time was>40-90 s at 22℃,the egg survival rate of mice with>0.1-0.5μl frozen fluid accumulation was higher than that of>0.5-1.0μl,and the difference was statistically significant(P<0.05).Conclusion The best freezing effect and the highest survival rate of mouse eggs with radiative crown should be selected at 22℃ambient temperature,40-90 s operation time,0.1-0.5μl frozen liquid.

关 键 词：小鼠卵子 双层麦管法 玻璃化冷冻 复苏 存活率 

分 类 号：R71[医药卫生—妇产科学]