Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of São Francisco Valley, Petrolina, PE 56300-990, Brazil.
Health Center Science, State University of Ceará, 60714-903 Fortaleza, CE, Brazil.
Domest Anim Endocrinol. 2019 Apr;67:71-79. doi: 10.1016/j.domaniend.2018.12.003. Epub 2018 Dec 18.
This study evaluated the in vitro development and maturation of ovine oocytes from secondary follicles cultured in serum-free medium containing fixed or sequential concentrations of recombinant human FSH (rhFSH). Follicles were cultured in α-MEM alone or with constant (500, 750, or 1,000 ng/mL) or sequential concentrations of rhFSH (seq. 1: day 6 = 500; day 12 = 750; day 18 = 1,000 ng/mL and seq. 2: day 6 = 100; day 12 = 500; day 18 = 1,000 ng/mL). At the end of the experiment, follicular survival was higher (P < 0.05) in 750 ng/mL rhFSH than the control and 1,000 ng/mL rhFSH. As early as day 6 of culture, antral cavity formation was observed in all treatments. Follicular diameter increased progressively and significantly in all treatments throughout 18 d of culture. Furthermore, addition of rhFSH to the medium promoted a significant increase in the percentage of fully grown oocytes in all treatments compared to α-MEM. Mitochondrial activity was higher in rhFSH treatments than in the control, except in rhFSH seq. 2 (P < 0.05). Maturation rates increased in oocytes from intact follicles cultured in 750 ng/mL rhFSH compared to the control (P < 0.05). In conclusion, rhFSH at 750 ng/mL maintained the survival of secondary follicles cultured in serum-free medium, improved oocyte growth, mitochondrial activity, and oocyte maturation.
本研究评估了在含固定或连续浓度重组人促卵泡激素（rhFSH）的无血清培养基中培养的来自次级卵泡的绵羊卵母细胞的体外发育和成熟。卵泡在α-MEM 中单独培养或与恒定（500、750 或 1000ng/mL）或连续浓度 rhFSH（seq.1：第 6 天=500；第 12 天=750；第 18 天=1000ng/mL 和 seq.2：第 6 天=100；第 12 天=500；第 18 天=1000ng/mL）培养。实验结束时，750ng/mL rhFSH 组的卵泡存活率（P < 0.05）高于对照组和 1000ng/mL rhFSH 组。早在培养的第 6 天，所有处理组均观察到腔前卵泡的形成。在整个 18 天的培养过程中，卵泡直径在所有处理组中均逐渐显著增加。此外，与α-MEM 相比，在培养基中添加 rhFSH 可显著增加所有处理组中完全成熟卵母细胞的比例。与对照组相比，rhFSH 处理组的线粒体活性更高，除了 rhFSH seq.2（P < 0.05）。与对照组相比，在 750ng/mL rhFSH 培养的完整卵泡中，卵母细胞的成熟率增加（P < 0.05）。总之，750ng/mL rhFSH 维持了无血清培养基中次级卵泡的存活，提高了卵母细胞的生长、线粒体活性和卵母细胞的成熟。
