Follicle Biology Laboratory (FOBI), Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel (VUB), Brussels, Belgium.
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre (RS), Brazil.
Biol Reprod. 2021 Apr 1;104(4):902-913. doi: 10.1093/biolre/ioab008.
In vitro maturation (IVM) is an assisted reproduction technique with reduced hormone-related side-effects. Several attempts to implement IVM in routine practice have failed, primarily due to its relatively low efficiency compared with conventional in vitro fertilization (IVF). Recently, capacitation (CAPA)-IVM-a novel two-step IVM method-has improved the embryology outcomes through synchronizing the oocyte nuclear and cytoplasmic maturation. However, the efficiency gap between CAPA-IVM and conventional IVF is still noticeable especially in the numerical production of good quality embryos. Considering the importance of glucose for oocyte competence, its metabolization is studied within both in vivo and CAPA-IVM matured mouse cumulus-oocyte-complexes (COCs) through direct measurements in both cellular compartments, from transcriptional and translational perspectives, to reveal metabolic shortcomings within the CAPA-IVM COCs. These results confirmed that within in vivo COC, cumulus cells (CCs) are highly glycolytic, whereas oocytes, with low glycolytic activity, are deviating their glucose towards pentose phosphate pathway. No significant differences were observed in the CAPA-IVM oocytes compared with their in vivo counterparts. However, their CCs exhibited a precocious increase of glycolytic activity during the pre-maturation culture step and activity was decreased during the IVM step. Here, specific alterations in mouse COC glucose metabolism due to CAPA-IVM culture were characterized using direct measurements for the first time. Present data show that, while CAPA-IVM CCs are able to utilize glucose, their ability to support oocytes during final maturation is impaired. Future CAPA-IVM optimization strategies could focus on adjusting culture media energy substrate concentrations and/or implementing co-culture strategies.
体外成熟 (IVM) 是一种辅助生殖技术,具有减少激素相关副作用的优点。尽管人们多次尝试将 IVM 应用于常规实践,但都以失败告终,主要是因为其效率相对较低,与传统的体外受精 (IVF) 相比。最近,一种新型两步法 IVM 技术——获能 (CAPA)-IVM,通过同步卵母细胞核和细胞质成熟,改善了胚胎学结局。然而,CAPA-IVM 与传统 IVF 之间的效率差距仍然很明显,尤其是在优质胚胎的数量方面。考虑到葡萄糖对卵母细胞活力的重要性,我们通过直接在细胞内进行测量,从转录和翻译的角度研究了其在体内和 CAPA-IVM 成熟的小鼠卵丘-卵母细胞复合物 (COC) 中的代谢情况,以揭示 CAPA-IVM COC 中的代谢缺陷。这些结果证实,在体内 COC 中,卵丘细胞 (CC) 具有高度的糖酵解活性,而卵母细胞的糖酵解活性较低,会将葡萄糖转向戊糖磷酸途径。与体内 COC 相比,CAPA-IVM 卵母细胞中未观察到明显差异。然而,它们的 CC 在预成熟培养阶段表现出糖酵解活性的提前增加,而在 IVM 阶段则减少。在这里,我们首次使用直接测量法对 CAPA-IVM 培养导致的小鼠 COC 葡萄糖代谢的特定变化进行了特征描述。目前的数据表明,虽然 CAPA-IVM CC 能够利用葡萄糖,但它们在最终成熟过程中支持卵母细胞的能力受损。未来的 CAPA-IVM 优化策略可以集中在调整培养基的能量底物浓度和/或实施共培养策略上。