激光打孔技术用于冷冻精子复苏后体外受精的研究

doi:10.3969/j.issn.1674-5817.2012.03.007

摘要/Abstract

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% vs 28.6%, 77.4% vs 52.3%,40.7% vs 12.7%,56.5% vs 14.6%,65% vs 6.5%; 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% vs 50.8%,74.3% vs 35.6%,75.7% vs 32.7%,79.5% vs 48.9%,59.5% vs 46.3%,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。

关键词: 小鼠, 精子冷冻, 卵子激光打孔

Abstract: 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.

Key words: mice, sperm cryopreservation, laser-punched oocytes

中图分类号:

Q95-33

刘丽均, 朱权凤, 郁丽丽, 张屹达, 施美莲. 激光打孔技术用于冷冻精子复苏后体外受精的研究[J]. 实验动物与比较医学, 2012, 32(3): 203-208.

LIU Li-jun, ZHU Quan-feng, YU Li-li, ZHANG Yi-da, SHI Mei-lian. Using Laser-Punched System on IVF of Fresh and Cryopreservated and Post-Thawed Sperm[J]. Laboratory Animal and Comparative Medicine, 2012, 32(3): 203-208.

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