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华通氏胶来源的条件培养基对体外不成熟卵母细胞成熟、胚胎发育和凋亡相关基因表达的影响。

The Effect of Wharton's Jelly-Derived Conditioned Medium on the In Vitro Maturation of Immature Oocytes, Embryo Development, and Genes Expression Involved in Apoptosis.

机构信息

Department of Biology, Faculty of Sciences, Arak University, Arak, Iran.

Department of Biology, Faculty of Sciences, University of Qom, Qom, Iran.

出版信息

Reprod Sci. 2024 Jan;31(1):190-198. doi: 10.1007/s43032-023-01345-2. Epub 2023 Sep 11.

Abstract

Oocyte cytoplasmic maturation is a crucial process during in vitro maturation (IVM), and finding an appropriate IVM medium that promotes oocyte competence is very critical in assisted reproductive technology (ART). The aim of this study was to investigate the effects of umbilical cord Wharton's jelly (WJ-MSCs)-derived conditioned media on the maturation of immature oocytes and their developmental potential in humans after IVM, as well as apoptotic gene expression. A total of 392 germinal vesicle (GV) oocytes were collected from 207 women aged 25-35 years and divided into two IVM groups: (1) control group, which was cultured in Cleav medium, and (2) experimental group, which was cultured in supernatants of umbilical cord Wharton's jelly as a conditioned medium (CM). First, WJ-MSCs were isolated, and their purity was analyzed. The immunophenotypes of WJ-MSCs were analyzed by flow cytometry. The quantitative expression of BCL2, BAX, and BAG1 in matured oocytes and embryos was evaluated through quantitative real-time polymerase chain reaction (qRT-PCR). Our findings showed that WJ-MSCs have a high proliferating capacity. The purity of the isolated cells was further validated by immunophenotyping, which revealed that their surface antigen expression had phenotypical properties similar to WJ-MSCs. When compared to CD34 and CD45 surface markers, the enlarged cells were positive for CD90, CD105, and CD44. There were significant differences in cytoplasmic maturation of oocytes and embryo quality between the two groups. The mRNA expression levels of BCL-2, BAG1, and BAX in matured oocytes and embryos were also significantly different between the two groups. Therefore, WJ-MSCs medium indicated potential efficacy in terms of ameliorating oocyte maturation and in promoting the development and genes expression of BAX, BCL-2, and BAG1.

摘要

卵母细胞胞质成熟是体外成熟(IVM)过程中的一个关键过程,寻找促进卵母细胞能力的合适 IVM 培养基在辅助生殖技术(ART)中非常关键。本研究旨在探讨脐带华通氏胶(WJ-MSCs)衍生的条件培养基对人类未成熟卵母细胞 IVM 后成熟和发育潜能的影响,以及凋亡基因表达。从 207 名年龄在 25-35 岁的女性中收集了 392 个生发泡(GV)卵母细胞,并将其分为两组 IVM 组:(1)对照组,在 Cleav 培养基中培养,(2)实验组,在脐带华通氏胶的上清液中培养作为条件培养基(CM)。首先,分离 WJ-MSCs,并分析其纯度。通过流式细胞术分析 WJ-MSCs 的免疫表型。通过定量实时聚合酶链反应(qRT-PCR)评估成熟卵母细胞和胚胎中 BCL2、BAX 和 BAG1 的定量表达。我们的研究结果表明,WJ-MSCs 具有高增殖能力。通过免疫表型进一步验证了分离细胞的纯度,结果表明其表面抗原表达具有与 WJ-MSCs 相似的表型特征。与 CD34 和 CD45 表面标志物相比,增大的细胞对 CD90、CD105 和 CD44 呈阳性。两组之间卵母细胞细胞质成熟和胚胎质量存在显著差异。成熟卵母细胞和胚胎中 BCL-2、BAG1 和 BAX 的 mRNA 表达水平也存在显著差异。因此,WJ-MSCs 培养基在改善卵母细胞成熟以及促进 BAX、BCL-2 和 BAG1 的发育和基因表达方面具有潜在的疗效。

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