获能培养液等因素对遗传工程小鼠冻融精子体外受精率的影响  

Effects of preincubation medium and other factors affecting in vitro fertilization rate of post-thawed genetically engineering mouse spermatozoa

作　　者：左琴[1] 范涛[1] 张翠平 杨文东 王劲松[1] 范昌发[1] 刘佐民[1] 贺争鸣[1] 李保文[1] 

机构地区：[1]中国食品药品检定研究院,国家啮齿类实验动物种子中心,北京100050 [2]北京百奥赛图基因生物技术有限公司,北京100076

出　　处：《中国比较医学杂志》2015年第1期45-49,共5页Chinese Journal of Comparative Medicine

基　　金：国家科技支撑计划(2013BAK11B00)

摘　　要：目的探讨遗传工程小鼠精子冷冻、复苏及体外受精率的效果,建立简便、经济的遗传工程小鼠保种体系。方法采用精子冷冻、体外受精和胚胎移植技术,比较了精子获能培养液、雄鼠周龄及精子冻存液等因素对于遗传工程小鼠精子冷冻保存的影响。结果用CPA精子冻存液冷冻保存精子,精子复苏后用PM精子培养液获能培养,体外受精率在82.49%-91.43%,而HTF精子培养液的体外受精率为14.46%-27.38%,同品系间差异极显著（P〈0.01）;10-35周龄的雄鼠精子均能成功冷冻、受精,移植后产仔;使用R18S3,CPM,CPA三种不同精子冻存液冷冻遗传工程小鼠精子,复苏后采用PM精子培养液体外受精,受精率分别是75.85%、88.89%和94.27%,移植后得到阳性小鼠;体外受精后的胚胎成功冷冻保存,移植后产仔。结论采用CPA精子冻存液冷冻保存精子,精子复苏后用PM精子培养液体外受精,能够更有效地保存遗传工程小鼠。Objective To discuss the effect of in vitro fertilization（ IVF） and mouse sperm cryopreservation,to establish a simple and economic frozen system for the genetically engineering mice preservation. Methods Sperm from genetically engineering mice were cryopreserved,IVF was performed using post-thawed sperm,then embryo transfer,to compare the effects of cryopreservation medium、age of male mice and sperm preincubation medium. Results Using CPA as sperm cryopreservation medium,when PM was used thawed-sperm preincubation in IVF,the fertility rates were from 82. 49%to 91. 43%,when HTF was used thawed-sperm preincubation in IVF,the fertility rates were from14. 46% to 27. 38%,therewas a signification difference between PM and HTF sperm preincubation medium; 10 to 35 weeks male genetically engineering mice sperm were succeed cryopreservation,and positive mice were procreated after 2-cell embryos were transferred; R18S3、CPM and CPA was used to freeze sperm,the fertility rates were 75. 85% 、88. 89% to 94. 27%,positive mice were procreated after 2-cell embryos were transferred; 2-cell embryos after IVF were freezed,then thawed and positive mice were procreated after 2-cell embryos were transferred. Conclusion Using CPA as sperm cryopreservation medium,when PM was used thawed-sperm preincubation in IVF,genetically engineering mice sperm were succeed cryopreservation.

关 键 词：精子 冷冻保存 体外受精 胚胎移植 遗传工程小鼠 

分 类 号：R332[医药卫生—人体生理学]