Advancing fertility preservation in prepubertal mice: Efficacy of ovarian tissue culture and in vitro growth in mature oocyte development

IF 1.6 4区医学 Q3 OBSTETRICS & GYNECOLOGY

Journal of Obstetrics and Gynaecology Research Pub Date : 2024-08-18 DOI:10.1111/jog.16057

Yuekun Chen, Yu Wakimoto, Mizuho Yano, Kohei Nakagawa, Akiko Hasegawa, Hiroaki Shibahara

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引用次数: 0

Abstract

Aim

This study aimed to evaluate the ovarian tissue culture and in vitro follicle growth as safer alternatives to cryopreservation for generating in vitro fertilization (IVF)-ready mature oocytes from prepubertal mice without the risk of cancer cell contamination.

Methods

Ovaries from prepubertal B6D2F1 mice were cultured in α-minimum essential medium supplemented with an estrogen receptor antagonist, ICI 182780. Culture duration was investigated to identify the optimal timeframe for follicle growth and oocyte maturation. Follicles were isolated mechanically or using 1 mg/mL collagenase and cultured in Matrigel matrix or polyvinylpyrrolidone. Oocyte development at metaphase II was induced by in vitro maturation, followed by IVF.

Results

The optimal culture duration was 2–4 days, and tissues cultured beyond this period showed significant follicular degeneration. ICI 182780 supplementation resulted in the recovery of 20.5 follicles per ovary compared with 9.5 follicles in non-supplemented cultures (p < 0.05). Of the 452 isolated follicles, 237 (52.4%) showed growth, 150 (33.2%) underwent germinal vesicle breakdown, and 18 (4.0%) reached metaphase II. However, none of the metaphase II oocytes were successfully fertilized after IVF. Matrigel demonstrated a significantly higher in vitro maturation rate compared with polyvinylpyrrolidone in a comparative analysis of culture matrices (p < 0.001).

Conclusions

This study highlighted ovarian tissue culture and in vitro growth as effective strategies for producing mature oocytes from prepubertal mice. Further studies are required to overcome fertilization hurdles and understand the mechanisms that improve post-IVF embryo viability.

Abstract Image

查看原文本刊更多论文

推进青春期前小鼠的生育能力保存：卵巢组织培养和成熟卵母细胞体外生长的功效。

目的：本研究旨在评估卵巢组织培养和体外卵泡生长作为冷冻保存的更安全替代方法，是否可用于从青春期前小鼠体内产生体外受精（IVF）准备就绪的成熟卵母细胞，且无癌细胞污染的风险：方法：在补充了雌激素受体拮抗剂 ICI 182780 的 α-最基本培养基中培养青春期前 B6D2F1 小鼠的卵巢。对培养持续时间进行了研究，以确定卵泡生长和卵母细胞成熟的最佳时间框架。用机械方法或 1 mg/mL 胶原酶分离卵泡，并在 Matrigel 基质或聚乙烯吡咯烷酮中培养。通过体外成熟诱导卵母细胞发育至分裂期 II，然后进行体外受精：结果：最佳培养时间为 2-4 天，超过这一期限的组织会出现明显的卵泡退化。补充 ICI 182780 后，每个卵巢恢复了 20.5 个卵泡，而未补充 ICI 182780 的培养物中仅恢复了 9.5 个卵泡（p 结论：ICI 182780 对卵巢组织培养和试管受精具有重要意义：本研究强调了卵巢组织培养和体外生长是生产青春期前小鼠成熟卵母细胞的有效策略。要克服受精障碍并了解提高体外受精后胚胎存活率的机制，还需要进一步研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Obstetrics and Gynaecology Research 医学-妇产科学

CiteScore

3.10

自引率

0.00%

发文量

376

审稿时长

3-6 weeks

期刊介绍： The Journal of Obstetrics and Gynaecology Research is the official Journal of the Asia and Oceania Federation of Obstetrics and Gynecology and of the Japan Society of Obstetrics and Gynecology, and aims to provide a medium for the publication of articles in the fields of obstetrics and gynecology. The Journal publishes original research articles, case reports, review articles and letters to the editor. The Journal will give publication priority to original research articles over case reports. Accepted papers become the exclusive licence of the Journal. Manuscripts are peer reviewed by at least two referees and/or Associate Editors expert in the field of the submitted paper.