建立遗传工程小鼠的冷冻保种体系.doc

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 左琴，，，贺争鸣*，* (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 【摘要目的简便、经济的方法 采用，结果P0.01）；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 【关键词；；； 【中图分类号】R - 33 【文献标识码】A【文章编号】 ZUO Qin1, FAN Tao1 , ZHANG Cui-ping2, YANG Wen-dong2, WANG Jin-song1, FAN Chang-fa1, LIU Zuo-min1, HE Zheng-ming1*, LI Bao-wen1* (1. Laboratory Animal Resource Center, National Institutes Food and Drug Control, Beijing 100050; 2.Beijing Biocytogen Co., Ltd, Beijing 100076, China) [Abstract] 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%, there was 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. [Key words]Sperm; Cryopreservation; In