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牛卵母细胞在卵丘-卵母细胞复合体中的体外生长及卵泡刺激素对卵母细胞生长的影响。

In vitro growth of bovine oocytes in oocyte-cumulus cell complexes and the effect of follicle stimulating hormone on the growth of oocytes.

机构信息

Laboratory of Developmental Biotechnology, Graduate School of Agricultural Science, Kobe University, Kobe 657-8501, Japan.

出版信息

J Reprod Dev. 2021 Feb 15;67(1):5-13. doi: 10.1262/jrd.2020-102. Epub 2020 Oct 30.

Abstract

Several successful in vitro culture experiments have used oocyte-cumulus cell-mural granulosa cell complexes (OCGCs) from early antral follicles (0.5-0.7 mm) for the growth of bovine oocytes. However, in studies related to in vitro oocyte maturation and in vitro embryo production, oocyte-cumulus cell complexes (OCCs) that have no mural granulosa cells have been widely used instead of OCGCs. The purpose of this study was to determine whether cumulus cells alone support oocyte growth. First, OCCs and OCGCs were cultured in vitro for 14 days to compare the integrity of the complexes as well as antrum formation. After 14 days, the diameter and meiotic competence of oocytes in OCCs and OCGCs were examined. Oocytes in OCCs grew fully and acquired meiotic competence similar to OCGCs, whereas antrum formation occurred later in OCCs as compared to OCGCs. Subsequently, the effects of follicle stimulating hormone (FSH) on in vitro growth of OCCs were examined for 14 days. When FSH was added to the culture medium, OCCs formed antrum-like structures one day earlier than those cultured without FSH. Oocytes cultured with 1 mIU/ml FSH grew fully and acquired meiotic competence. In contrast, when oocytes were cultured in media containing high concentrations of FSH, some of the OCCs collapsed and the number of degenerated oocytes increased. In conclusion, bovine oocytes in OCCs grow and acquire meiotic competence similar to OCGCs and, 1 mIU/ml FSH supports the development of OCCs and oocyte growth as observed in our culture system.

摘要

几项成功的体外培养实验使用来自早期窦前卵泡(0.5-0.7mm)的卵丘-卵母细胞-颗粒细胞复合体(OCGCs)来生长牛卵母细胞。然而,在与体外卵母细胞成熟和体外胚胎生产相关的研究中,广泛使用没有壁颗粒细胞的卵丘-卵母细胞复合体(OCCs)代替 OCGCs。本研究旨在确定是否只有卵丘细胞支持卵母细胞生长。首先,将 OCCs 和 OCGCs 在体外培养 14 天,以比较复合体的完整性和窦腔形成。14 天后,检查 OCCs 和 OCGCs 中卵母细胞的直径和减数分裂能力。OCCs 中的卵母细胞完全生长并获得与 OCGCs 相似的减数分裂能力,而与 OCGCs 相比,OCCs 中的窦腔形成较晚。随后,研究了卵泡刺激素(FSH)对 OCCs 体外生长的影响,培养 14 天。当 FSH 添加到培养基中时,OCCs 形成窦状结构的时间比不添加 FSH 的培养基早一天。在含有 1mIU/ml FSH 的培养基中培养的卵母细胞完全生长并获得减数分裂能力。相比之下,当卵母细胞在含有高浓度 FSH 的培养基中培养时,一些 OCCs 塌陷,退化卵母细胞的数量增加。总之,在 OCCs 中的牛卵母细胞生长并获得与 OCGCs 相似的减数分裂能力,并且 1mIU/ml FSH 支持我们的培养系统中观察到的 OCCs 和卵母细胞生长的发育。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b5ec/7902213/a2f0da782c92/jrd-67-005-g001.jpg

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