马卵母细胞体外成熟的研究进展

Research Progress of In Vitro Maturation in Horse Oocytes

随着全球马产业的发展,马发挥的经济价值越来越大。辅助生殖技术有利于发挥优良马匹的潜在价值。马卵母细胞体外成熟(IVM)是辅助生殖技术重要的组成部分,卵母细胞的获取是体外成熟的前提,切刮法能从离体卵巢中获得较多的马卵母细胞,而活体采卵技术(OPU)则能持续地获得卵母细胞,并能较好的保存马卵母细胞的发育能力。扩张型卵母细胞的成熟率高于紧密型卵母细胞,母马的年龄会影响到其卵母细胞的质量。马卵母细胞体外存放较长时间不会影响其发育能力,现在已有较为成熟的体系能使马卵母细胞在体外保存24 h以上而不影响其成熟率。在马卵母细胞成熟体系中常用的基础培养液是M199,添加胎牛血清(FBS)、促卵泡素(FSH)、促黄体生成素(LH)、胰岛素样生长因子-1(IGF-1)等物质能显著提高成熟率,常用培养环境为38~39℃,5%CO2饱和湿度下培养,培养时间30 h。成熟的卵母细胞有扩张的卵丘细胞和极体,且成熟的卵母细胞的细胞骨架及微管结构也会发生变化。本文针对马卵母细胞的采集和体外成熟培养的相关研究进行总结,重点阐述了不同采集技术的回收率以及影响马卵母细胞体外成熟率的关键因素,以期对今后马卵母细胞体外成熟的进一步研究及后期体外受精技术的发展提供借鉴与参考。

With the development of the global horse industry,the economic value of horses is growing.Assisted reproductive technology is beneficial to dig out the potential value of excellent equine,and in vitro maturation(IVM)of horse oocytes is an important part of assisted reproductive technology.Oocyte recovery is a preparation for in vitro maturation,for the isolated ovary,more oocytes can be obtained by using the scraping method.To obtain oocytes continuously for a long time and to protect the development ability of oocytes,ovum pick up(OPU)is widely used in mare.The maturation rate of expanded oocytes is higher than that of compact oocytes,and the age of the mares affects the quality of their oocytes.Horse oocytes stored in vitro for a long time will not affect their developmental ability,there are mature systems that can keep horse oocytes in vitro for more than 24 hours without affecting their maturation rate.The basic culture medium commonly used in horse oocyte maturation system is M199,and the addition of FBS,FSH,LH,IGF-1 and other substances can significantly improve the maturation rate,and the commonly used culture environment is 38-39℃,5%CO2 and saturated humidity incubate for 30 hours.Mature oocytes have dilated cumulus cells and polar body,and the cytoskeleton and microtubule structure of oocytes will be changed after maturation.This review summarized the research on the collection and in vitro maturation of equine oocytes,focusing on the recovery rate of different collection techniques and some key factors affecting the in vitro maturation rate of equine oocytes,in order to provide some knowledge and experience for further research of IVM and in vitro fertilization(IVF)in the future.