Douet Cécile, Parodi Olivia, Martino Nicola Antonio, Lacalandra Giovanni Michele, Nicassio Michele, Reigner Fabrice, Deleuze Stefan, Dell'Aquila Maria Elena, Goudet Ghylène
PRC,INRA,CNRS,IFCE,Université de Tours,37380 Nouzilly,France.
Istituto Zooprofilattico Sperimentale per la Puglia e la Basilicata,Foggia,Italy.
Zygote. 2017 Oct;25(5):612-630. doi: 10.1017/S096719941700048X. Epub 2017 Sep 20.
Most wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) technique for equine embryo production is available. The development of culture conditions able to mimic the maturation of the oocyte in preovulatory follicular fluid (pFF) and the post-maturation in oviductal fluid (OF) may improve embryo production in vitro. Our aim was to analyse the effect of in vitro maturation in pFF and incubation in OF on in vitro maturation of equine oocytes, fertilization using conventional IVF or ICSI, and embryo development after culture in synthetic oviductal fluid (SOF) or DMEM-F12. Oocytes collected from slaughtered mares or by ovum pick up were matured in vitro in pFF or semi-synthetic maturation medium (MM). The in vitro maturation, fertilization and development rates were not statistically different between pFF and MM. After in vitro maturation, oocytes were incubated with or without OF. Post-maturation in OF did not significantly improve the fertilization and development rates. Thus, in our study, exposure to physiological fluids for oocyte maturation and post-maturation does not improve in vitro embryo production in the horse.
大多数野生马科动物和许多家养马品种都面临灭绝风险，因此迫切需要建立基因组资源库。胚胎冷冻保存能够保存雌雄双方的遗传物质，是恢复一个品种最快的方法。在马科动物中，体外胚胎生产可使每个周期产生多个胚胎。胞浆内单精子注射（ICSI）用于生产马胚胎，但它需要昂贵的设备和显微操作专业技术，而且囊胚发育率仍然较低。目前尚无用于马胚胎生产的传统体外受精（IVF）技术。开发能够模拟排卵前卵泡液（pFF）中卵母细胞成熟以及输卵管液（OF）中成熟后状态的培养条件，可能会提高体外胚胎生产效率。我们的目的是分析在pFF中体外成熟以及在OF中孵育对马卵母细胞体外成熟、使用传统IVF或ICSI受精以及在合成输卵管液（SOF）或DMEM-F12中培养后的胚胎发育的影响。从屠宰母马采集或通过采卵获得的卵母细胞在pFF或半合成成熟培养基（MM）中进行体外成熟培养。pFF和MM之间的体外成熟、受精和发育率在统计学上没有差异。体外成熟后，卵母细胞在有或无OF的情况下进行孵育。在OF中成熟后并没有显著提高受精和发育率。因此，在我们的研究中，使卵母细胞暴露于生理液体中进行成熟和成熟后培养并不能提高马的体外胚胎生产效率。
