Use of platelet-rich plasma on in vitro maturation during bovine embryo production.

IF 1.6 4区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Animal Reproduction Pub Date : 2024-03-22 eCollection Date: 2024-01-01 DOI:10.1590/1984-3143-AR2023-0107
Eduardo Baia de Souza, Diego Dubeibe Marin, Anelise Sarges Ramos, Bruno Porpino Homobono, Priscilla do Carmo de Azevedo Ramos, Vanessa Cunha de Brito, Gabriela Santos da Cruz, Nathalia Nogueira da Costa, Marcela da Silva Cordeiro, Simone do Socorro Damasceno Santos
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Abstract

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.

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在牛胚胎生产过程中使用富血小板血浆促进体外成熟。
胚胎体外培养(IVP)要取得成功,必须考虑的一个重要方面是在这项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配子在卵母细胞成熟和精子发育过程中以及胚胎在初始细胞分裂过程中的代谢需求。大多数体外受精方案在培养基成分中加入血清,作为激素、蛋白质、生长因子和营养物质的来源。许多研究都建议用血小板丰富血浆(PRP)替代细胞培养中的胎儿血清,尤其是干细胞。因此,本研究的目的是评估在体外生产牛胚胎的卵母细胞成熟过程中使用富血小板血浆替代胎牛血清(FBS)的可能性。在体外成熟(IVM)过程中,将积液-卵母细胞复合体(COCs)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积液-卵母细胞复合体受精,并将所产生的子代细胞培养七天。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。此外,还通过分析与细胞应激有关的重要蛋白质 HSP70 的基因表达来评估成熟 COC 的质量。结果表明,各实验组在胚胎生产率方面没有显著差异,无论是在最初的分裂阶段还是囊胚形成阶段(G4 组除外,该组在第 7 天表现出较低的囊胚形成率,正如预期的那样)。这表明,在胚胎的 IVP 中,PRP 可作为 SFB 的一种经济有效的替代品。
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来源期刊
Animal Reproduction
Animal Reproduction AGRICULTURE, DAIRY & ANIMAL SCIENCE-
CiteScore
2.30
自引率
11.80%
发文量
49
审稿时长
70 days
期刊介绍: Animal Reproduction (AR) publishes original scientific papers and invited literature reviews, in the form of Basic Research, Biotechnology, Applied Research and Review Articles, with the goal of contributing to a better understanding of phenomena related to animal reproduction. The scope of the journal applies to students, researchers and practitioners in the fields of veterinary, biology and animal science, also being of interest to practitioners of human medicine. Animal Reproduction Journal is the official organ of the Brazilian College of Animal Reproduction in Brazil.
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