Facultad de Zootecnia y Ecología, Universidad Autónoma de Chihuahua. Chihuahua, Chihuahua, Mexico.
Facultad de Zootecnia y Ecología, Universidad Autónoma de Chihuahua. Chihuahua, Chihuahua, Mexico.
Theriogenology. 2020 Apr 1;146:1-13. doi: 10.1016/j.theriogenology.2020.01.049. Epub 2020 Jan 29.
Starting in vitro fertilization process with competent oocytes that may endure first cellular divisions is a critical step for obtaining an embryo. To obtain in vitro competent oocytes, culture conditions should emulate the in vivo microenvironment as close as possible. With the aim of improving the in vitro culture medium, the present study evaluated the IMD/ADM2 peptide as a factor that promotes oocyte competence and improves embryo quality in bovine systems. The culture supplemented with 153 μg/mL of IMD/ADM2 was correlated with the production of healthy oocytes in metaphase II (MII) stage in compacted cumulus-oocyte complexes (COC) with a decrease of BAX/BCL-2 to mRNA ratio and a reduction of late apoptosis by TUNEL in MII oocytes. In addition to this, treatment with IMD/ADM2 caused cAMPi level to be constant over time, and the cAMPi level kept increasing until 6 h. COC supplementation with 153 μg/mL of IMD/ADM2 increased the blastocyst production rate two-fold in comparison with control conditions. Only embryos from COC treatment with this peptide were capable of developing blastocysts in stage-6 grade I; compared with the control culture, it was the treatment with the greater number of blastocysts stage-5; these are characteristics of good quality blastocysts.
从具有能够耐受第一次细胞分裂的成熟卵母细胞开始体外受精过程是获得胚胎的关键步骤。为了获得体外成熟的卵母细胞,培养条件应尽可能接近体内微环境。本研究旨在通过添加 IMD/ADM2 肽来改善体外培养体系,以提高牛卵母细胞的成熟能力并改善胚胎质量。结果显示,在 IMD/ADM2 浓度为 153μg/mL 的培养基中培养的卵母细胞在致密的卵丘-卵母细胞复合物(COC)中处于中期 II(MII)阶段,且 BAX/BCL-2 比值降低,晚期凋亡减少,同时,COC 中添加 IMD/ADM2 可使 cAMPi 水平随时间保持恒定,并在 6 小时内持续增加。与对照组相比,添加 153μg/mL IMD/ADM2 的 COC 使囊胚的生成率增加了一倍。只有经 IMD/ADM2 处理的 COC 才能发育成阶段 6 级 I 的囊胚,与对照组相比,这是处理后的囊胚数量更多,达到了 5 阶段;这些都是优质囊胚的特征。