Ahmadi Ebrahim, Shirazi Abolfazl, Shams-Esfandabadi Naser, Nazari Hassan
Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.
Reprod Domest Anim. 2019 Mar;54(3):595-603. doi: 10.1111/rda.13402. Epub 2019 Feb 11.
Despite the numerous potential applications of oocyte cryopreservation, the poor success rate has limited its practical applications. In livestock, particularly in ovine, the oocytes have low developmental competence following vitrification/warming process. Considering the occurrence of osmotic and oxidative stresses during the vitrification/warming process, the application of antioxidants and osmolytes may improve the developmental competence of vitrified/warmed oocytes. In the present study, we aimed to evaluate the effects of the addition of ascorbic acid (AA) and N-acetyl cysteine (NAC) as antioxidants and glycine as an organic osmolyte either to the vitrification/warming solutions (VWS) or to the IVM medium on the developmental competence of vitrified/warmed ovine germinal vesicle stage oocytes. The survival rate in the vitrified groups was significantly lower than fresh ones. In vitrified/warmed oocytes, there was no significant difference in survival rate between supplemented and non-supplemented groups. The addition of AA and/or NAC to the VWS or IVM medium and adding glycine to the IVM medium reduced the proportion of apoptotic oocytes and fragmented embryos, which was reflected as an increase in the proportions of metaphase II stage oocytes and blastocyst production. The best result was achieved by supplementing the IVM medium with NAC. In our study condition, antioxidants and glycine could improve the developmental competence of vitrified/warmed ovine immature oocytes, especially when added during IVM.
尽管卵母细胞冷冻保存有众多潜在应用,但成功率低限制了其实际应用。在畜牧业中,尤其是绵羊,卵母细胞在玻璃化/复温过程后发育能力较低。考虑到玻璃化/复温过程中会出现渗透应激和氧化应激,添加抗氧化剂和渗透压调节剂可能会提高玻璃化/复温后卵母细胞的发育能力。在本研究中,我们旨在评估向玻璃化/复温溶液(VWS)或体外成熟(IVM)培养基中添加抗氧化剂抗坏血酸(AA)和N-乙酰半胱氨酸(NAC)以及有机渗透压调节剂甘氨酸对玻璃化/复温后的绵羊生发泡期卵母细胞发育能力的影响。玻璃化组的存活率显著低于新鲜组。在玻璃化/复温的卵母细胞中,添加组和未添加组的存活率没有显著差异。向VWS或IVM培养基中添加AA和/或NAC以及向IVM培养基中添加甘氨酸可降低凋亡卵母细胞和碎片化胚胎的比例,这表现为中期II期卵母细胞比例和囊胚生成率的增加。通过向IVM培养基中添加NAC取得了最佳结果。在我们的研究条件下,抗氧化剂和甘氨酸可提高玻璃化/复温后的绵羊未成熟卵母细胞的发育能力,尤其是在IVM期间添加时。