激光打孔技术用于冷冻精子复苏后体外受精的研究  

Using Laser-Punched System on IVF of Fresh and Cryopreservated and Post-Thawed Sperm

作　　者：刘丽均 朱权凤 郁丽丽 张屹达 施美莲 

机构地区：[1]上海斯莱克实验动物有限责任公司,上海201615

出　　处：《实验动物与比较医学》2012年第3期203-208,共6页Laboratory Animal and Comparative Medicine

基　　金：上海市科委实验动物专项课题（10140900600）

摘　　要：目的比较精子冷冻前后体外受精率的变化，以及卵子经过激光打孔后，与新鲜精子及冷冻复苏精子体外受精，比较受精率的变化。方法冷冻2个近交系小鼠和5个品系的基因工程小鼠的精子，比较新鲜精子和冷冻复苏精子与C57BL／6J小鼠卵子做体外受精的结果：然后对C57BL／6J卵子做激光打孔，再与新鲜及冷冻复苏精子体外受精结果进行比较。结果（1）新鲜C57BL／6J精子受精率为50．4％，C57BL／6N的受精率为48．6％，远高于冷冻精子复苏后的受精率（17．7％和15．4％）；卵子打孔后，再与C57BL／6J及C57BL／6N的新鲜精子或冷冻复苏精子受精，受精率分别为64．6％，59．7％和60．4％，48．3％，冷冻精子复苏后的体外受精率提高显著；（2）5个基因工程小鼠新鲜精子和冷冻精子复苏后与C57BL／6J卵子的受精率分别为：91．5％VS28．6％，77．4％1）S52．3％，40．7％VS12．7％，56．5％VS14．6％，65％VS6．5％；这5个品系的小鼠新鲜精子和冷冻精子复苏后与C57BL／6J打孔卵子的受精率分别为75％VS50．8％，74．3％FS35．6％，75．7％VS32．7％，79．5％VS48．9％，59．5％VS46．3％，冷冻精子复苏后的受精率较之不打孔卵子，提高显著。结论激光打孔技术用于冷冻复苏精子的辅助体外受精，可以显著提高冷冻精子复苏后的受精率，使得精子保种更加经济有效。Objective The IVF rate of the fresh sperm and the post-thawed sperm about C57BL/6J mice were compared, and then C57BL/6J oocytes were laser-punched, and these oocytes were IVF with fresh and post-thawed sperm, their IVF rate were investigated. Method Two inbred mice strains of （C57BL/6J, C57BL/6N） and 5 strains of genetic engineering mice （MKP-5, Sir-168, C57BL/6RLR 3-/-, C57BL/6 TRL 2KO, C57BL/6 TRL 4KO） were chosen to freeze sperms. Firstly, the IVF rate of fresh sperm and post-thawed sperm with normal C57BL/6J oocytes were compared. Secondly, C57BL/6J oocytes were laser-punched, and then the IVF rate of the fresh sperm and post-thawed sperm with these oocytes were compared. Result （1） The IVF rate of fresh sperm and post-thawed sperm of C57BL/ 6J, C57BL/6N were 50.4% vs 17.7%, 48.6% vs 15.4%; when the oocytes were laser-punched, the IVF rate were 64.6% vs 59.7%, 60.4% vs 15.4%; （2） the IVF rate of fresh and frozen sperm of 5 genetic engineering mice were 91.5% vs 28.6%, 77.4% vs 52.3%, 40.7% vs 12.7%, 56.5% vs 14.6%, 65% vs 6. 5%, when those sperm were fertilized with punched C57BL/6J oocytes, the result were 75% vs 50.7%, 74.3% vs 35.6%, 75.7% vs 32.7%, 79.5% vs 48.9%, 59.5% vs 46.3%. Conclusion When oocytes were laser-punched, the IVFrate of post-thawed sperm were increased significantly, it is showed that laser-punched system was a useful and economical way to increase the IVF rate of mice frozen sperm.

关 键 词：小鼠 精子冷冻 卵子激光打孔 

分 类 号：R714.8[医药卫生—妇产科学] R-332[医药卫生—临床医学]