Souza-Fabjan Joanna Maria Gonçalves, Locatelli Yann, Duffard Nicolas, Corbin Emilie, Batista Ribrio Ivan Tavares Pereira, de Figueirêdo Freitas Vicente José, Beckers Jean-François, Mermillod Pascal
INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France; Laboratory of Physiology and Control of Reproduction, Veterinary School, State University of Ceará, Fortaleza, CE, Brazil.
INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France; Museum National d'Histoire Naturelle, Réserve de la Haute Touche, Obterre, France.
Theriogenology. 2016 Nov;86(8):1989-98. doi: 10.1016/j.theriogenology.2016.06.021. Epub 2016 Jun 29.
Although cumulus cells are essential for efficient oocyte maturation, the establishment of protocols that support IVD of embryos obtained from denuded oocytes (DOCs) is important for optimizing the use of reproductive biotechnologies. Thus, this study aimed to establish a protocol for IVD of goat DOC using different strategies of IVM and methods of oocyte activation. Four experiments were performed. Similar developmental competence of slaughterhouse DOC was obtained, regardless of maturation media (complex, semidefined or simplified). However, the ability to reach the blastocyst stage was affected by the activation method. Denuded oocytes subjected to parthenogenetic activation had greater (P < 0.05) development capacity, compared with those undergoing IVF with average cleavage rate of 83% and 75%, blastocyst rate of 49% and 28%, and blastocysts in relation to the cleaved embryos of 59% and 38, respectively. In addition, the quality of embryos evaluated after vitrification/warming was similar between parthenogenetic activation and IVF. Finally, we demonstrated that the coculture of cumulus-oocyte complex (COC) with DOC increased the competence of DOC at a ratio of 1:1 and 1:9 (DOC:COC). We believe that presence of cumulus cells (CCs) is not essential to the meiotic maturation, if at the time of removal of the oocyte from follicular environment, they already acquired competence to development. However, when the oocytes still need to acquire competence, the presence of CC may significantly contribute in their developmental capacity acquisition during IVM. Thus, regardless of the source, these oocytes will require longer time in IVM, contrary to what happens in the absence of CC. In conclusion, although DOC had a lower developmental potential, especially after IVF, they were able to produce blastocysts and the coculture of DOC with COC increased this developmental capacity.
尽管卵丘细胞对于卵母细胞的高效成熟至关重要,但建立支持从裸卵(DOCs)获得的胚胎进行体外发育(IVD)的方案对于优化生殖生物技术的应用非常重要。因此,本研究旨在建立一种使用不同体外成熟(IVM)策略和卵母细胞激活方法的山羊DOC体外发育方案。进行了四项实验。无论成熟培养基(复杂、半限定或简化)如何,屠宰场DOC都具有相似的发育能力。然而,达到囊胚阶段的能力受激活方法的影响。与体外受精(IVF)的裸卵相比,孤雌激活的裸卵具有更高的(P < 0.05)发育能力,IVF的平均卵裂率分别为83%和75%,囊胚率分别为49%和28%,相对于卵裂胚胎的囊胚率分别为59%和38%。此外,玻璃化/复温后评估的胚胎质量在孤雌激活和IVF之间相似。最后,我们证明卵丘 - 卵母细胞复合体(COC)与DOC以1:1和1:9(DOC:COC)的比例共培养可提高DOC的发育能力。我们认为,如果在从卵泡环境中取出卵母细胞时它们已经获得了发育能力,那么卵丘细胞(CCs)的存在对于减数分裂成熟并非必不可少。然而,当卵母细胞仍需要获得发育能力时,CC的存在可能会在体外成熟期间对其发育能力的获得做出显著贡献。因此,无论来源如何,这些卵母细胞在体外成熟中都需要更长的时间,这与没有CC时的情况相反。总之,尽管DOC的发育潜力较低,尤其是在体外受精后,但它们能够产生囊胚,并且DOC与COC的共培养提高了这种发育能力。