de Souza Eduardo Baia, Marin Diego Dubeibe, Ramos Anelise Sarges, Homobono Bruno Porpino, Ramos Priscilla do Carmo de Azevedo, de Brito Vanessa Cunha, da Cruz Gabriela Santos, da Costa Nathalia Nogueira, Cordeiro Marcela da Silva, Santos Simone do Socorro Damasceno
Laboratório de Fertilização in Vitro, Instituto de Ciências Biológicas - ICB, Universidade Federal do Pará - UFPA, Belém, PA, Brasil.
Facultad de Medicina Veterinaria y Zootecnia, Universidad de Ciencias Aplicadas y Ambientales - UDCA, Bogotá, Colombia.
Anim Reprod. 2024 Mar 22;21(1):e20230107. doi: 10.1590/1984-3143-AR2023-0107. eCollection 2024.
One of the crucial aspects to be considered for successful in vitro production (IVP) of embryos is the composition of the various media used throughout the stages of this reproductive biotechnology. The cell culture media employed should fulfill the metabolic requirements of both gametes during oocyte maturation and sperm development, as well as the embryo during its initial cell divisions. Most IVP protocols incorporate blood serum into the media composition as a source of hormones, proteins, growth factors, and nutrients. Numerous studies have suggested Platelet-Rich Plasma (PRP) as a substitute for fetal sera in cell culture, particularly for stem cells. Therefore, the objective of this study is to assess the potential use of PRP as a replacement for fetal bovine serum (FBS) during oocyte maturation for in vitro production of bovine embryos. During in vitro maturation (IVM), cumulus-oocyte complexes (COCs) were allocated into the following experimental groups: Group G1 (IVM medium with 5% PRP); Group G2 (MIV medium with 5% PRP and 5% SFB); Group G3 (MIV medium with 5% SFB); and Group G4 (MIV medium without either PRP or SFB). Subsequently, the cumulus-oocyte complexes were fertilized with semen from a single bull, and the resulting zygotes were cultured for seven days. Cleavage and blastocyst formation rates were assessed on days 2 and 7 of embryonic development, respectively. The quality of matured COCs was also evaluated by analyzing the gene expression of HSP70, an important protein associated with cellular stress. The results demonstrated that there were no significant differences among the experimental groups in terms of embryo production rates, both in the initial cleavage stages and blastocyst formation (except for the G4 group, which exhibited a lower blastocyst formation rate on D7, as expected). This indicates that PRP could be a cost-effective alternative to SFB in the IVP of embryos.
胚胎体外生产(IVP)成功的关键因素之一是在这种生殖生物技术的各个阶段所使用的各种培养基的成分。所使用的细胞培养基应满足卵母细胞成熟和精子发育过程中配子的代谢需求,以及胚胎在其最初细胞分裂阶段的代谢需求。大多数IVP方案将血清纳入培养基成分中,作为激素、蛋白质、生长因子和营养物质的来源。许多研究表明富血小板血浆(PRP)可替代细胞培养中的胎牛血清,特别是用于干细胞培养。因此,本研究的目的是评估PRP在牛胚胎体外生产的卵母细胞成熟过程中替代胎牛血清(FBS)的潜在用途。在体外成熟(IVM)过程中,将卵丘-卵母细胞复合体(COC)分配到以下实验组:G1组(含5%PRP的IVM培养基);G2组(含5%PRP和5%SFB的MIV培养基);G3组(含5%SFB的MIV培养基);G4组(不含PRP或SFB的MIV培养基)。随后,用一头公牛的精液使卵丘-卵母细胞复合体受精,并将所得受精卵培养7天。分别在胚胎发育的第2天和第7天评估卵裂率和囊胚形成率。还通过分析与细胞应激相关的重要蛋白质HSP70的基因表达来评估成熟COC的质量。结果表明,在胚胎生产速率方面,各实验组在初始卵裂阶段和囊胚形成方面均无显著差异(除G4组外,如预期的那样,该组在第7天的囊胚形成率较低)。这表明在胚胎的IVP中,PRP可能是SFB的一种经济有效的替代品。