Optimization of vitrification methods for equine oocytes

IF 2.7 4区生物学 Q1 ANATOMY & MORPHOLOGY Tissue & cell Pub Date : 2024-11-19 DOI:10.1016/j.tice.2024.102632

Ming Du , Xinyu Li , Bayinnamula , Na Wang , Yuanyi Liu, Lei Zhang, Yiping Zhao, Manglai Dugarjaviin

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Abstract

An important method for preserving equine germplasm is the cryopreservation of equine oocytes. Due to its ease, rapidity and affordability, vitrification freezing has taken over as the primary method of horse oocyte cryopreservation. The vitrification cryoprotectants utilized in this investigation were Ethylene glycol (E), Dimethyl sulfoxide (D), Sucrose (S), and Ficoll (F). According to the oocyte volume alteration, the treatment time was 39 s in equilibrium solution ED10 (10 % EG + 10 % DMSO), 32 s in equilibrium solution ED15 (15 % EG + 15 % DMSO), while 20 s in equilibrium solution ED20 (20 % EG + 20 % DMSO). We prepared three kinds of cryosolutions EDFS30 (E15 %+D15 %+70 %FS), EDFS35 (E17.5 % + D17.5 % + 65 %FS), EDFS40 (E20 % + D20 % + 60 %FS) according to the proportion of protectant components. Among 27 freezing protocols, we selected protocol ED10 (39 s) + EDFS30 + 80 s which has the highest in vitro culture maturation rate of 19.3 % while protocol ED20 (20 s) + EDFS40 + 120 s is the worst. Apoptosis gene analysis revealed that BAX, BID, BOK, and TP53 expression was substantially higher in oocytes from the ED20 (20 s) + EDFS40 + 120 s group than in oocytes from the ED10 (39 s) + EDFS30 + 80 s and control groups (p<0.01). This study investigated several vitrification schemes for equine oocytes.

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优化马卵母细胞玻璃化方法。

马卵母细胞冷冻保存是保存马种质的一种重要方法。玻璃化冷冻因其简便、快速和经济实惠，已成为马卵母细胞冷冻保存的主要方法。本研究使用的玻璃化冷冻保护剂包括乙二醇（E）、二甲亚砜（D）、蔗糖（S）和Ficoll（F）。根据卵母细胞体积的变化，平衡溶液 ED10（10 % EG + 10 % DMSO）的处理时间为 39 秒，平衡溶液 ED15（15 % EG + 15 % DMSO）的处理时间为 32 秒，平衡溶液 ED20（20 % EG + 20 % DMSO）的处理时间为 20 秒。根据保护剂成分的比例，我们制备了 EDFS30（E15 %+D15 %+70 %FS）、EDFS35（E17.5 %+D17.5 %+65 %FS）、EDFS40（E20 %+D20 %+60 %FS）三种冷冻溶液。在27种冷冻方案中，我们选择了ED10（39 s）+ EDFS30 + 80 s方案，该方案的体外培养成熟率最高，为19.3%，而ED20（20 s）+ EDFS40 + 120 s方案的体外培养成熟率最差。凋亡基因分析表明，ED20（20 秒）+ EDFS40 + 120 秒组的卵母细胞中 BAX、BID、BOK 和 TP53 的表达量大大高于 ED10（39 秒）+ EDFS30 + 80 秒组和对照组（P＜0.05）。

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.