School of Biology, Damghan University, Damghan, Iran.
School of Biology, Damghan University, Damghan; Institute of Biological Sciences, Damghan University, Damghan, Iran.
Cryo Letters. 2021 Mar-Apr;42(2):67-72.
Cryopreservation can induce cellular, genomic, and epigenetic abnormalities.
To analyse the impact of ovarian vitrification on follicular development and its epigenetic effect on promoter methylation of Inhba and Inhbb in granulosa cells.
Mouse ovaries were divided into control, toxicity, and vitrified groups. The growth and development of follicles were examined. After in vitro culture of follicles, DNA was extracted from isolated granulosa cells and treated with sodium bisulfite. The promoter methylation of Inhba and Inhbb was analyzed by direct PCR sequencing.
Vitrification reduced the growth of follicles; however, antral cavity formation was not influenced negatively. Vitrification reduced the percentage of 5-methylcytosine in the Inhba promoter, while CpG sites in the promoter of Inhbb remained unmethylated.
Vitrification had adverse effects on follicle growth and the epigenetics of granulosa cells. The results of the current study show that vitrification methods of ovary need more improvement.
冷冻保存会导致细胞、基因组和表观遗传异常。
分析卵巢玻璃化对卵泡发育的影响及其对颗粒细胞中 Inhba 和 Inhbb 启动子甲基化的表观遗传效应。
将小鼠卵巢分为对照组、毒性组和玻璃化组。观察卵泡的生长和发育情况。体外培养卵泡后，从分离的颗粒细胞中提取 DNA 并进行亚硫酸氢盐处理。通过直接 PCR 测序分析 Inhba 和 Inhbb 启动子的甲基化情况。
玻璃化处理降低了卵泡的生长速度，但对腔前卵泡的形成没有负面影响。玻璃化处理降低了 Inhba 启动子中 5-甲基胞嘧啶的百分比，而 Inhbb 启动子中的 CpG 位点保持未甲基化状态。
玻璃化处理对卵泡生长和颗粒细胞的表观遗传有不良影响。本研究结果表明，卵巢玻璃化方法需要进一步改进。
