Widjiati Widjiati, Darsini Ninik, Hendrawan Viski Fitri, Taqwa Sultan Fadhilla, Shabira Zahra, Kurniawati Devia Yoanita
Department of Veterinary Anatomy, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia.
Department of Medical Biology, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.
Vet World. 2025 Jul;18(7):2127-2135. doi: 10.14202/vetworld.2025.2127-2135. Epub 2025 Jul 30.
Indonesia's indigenous Kacang goat population is in decline, posing a threat to food security and genetic diversity. maturation and cryopreservation techniques are key strategies for genetic conservation. However, heat shock stress during cryopreservation can compromise oocyte viability. This study evaluates the post-warming quality of Kacang goat oocytes exposed to different cryoprotectants. This study aimed to compare the efficacy of a modified cryoprotectant (30% ethylene glycol + 1M sucrose) with a commercial cryoprotectant in preserving post-warming oocyte quality, based on maturation rates and biomarker levels (heat shock protein 70 [HSP70], adenosine triphosphate [ATP], and glutathione [GSH]).
Oocytes were collected from goat ovaries and matured for 22 h. They were divided into three groups: Control (no vitrification), commercial cryoprotectant (T1), and modified cryoprotectant (T2). Post-warming quality was assessed using an enzyme-linked immunosorbent assay to quantify HSP70, ATP, and GSH levels. Statistical analysis included one-way analysis of variance and Pearson's correlation (p < 0.05).
Maturation rates were comparable across groups (control group [CG]: 84.3%, T1: 79.8%, T2: 77.2%; p > 0.05). HSP70 levels were significantly elevated in T2 compared to CG (p < 0.05). T2 also showed significantly higher ATP (52.13 ± 7.7 ng/mL) and GSH (1.27 ± 0.66 ng/mL) levels compared to T1 (ATP: 25.65 ± 1.63; GSH: 0.06 ± 0.01 ng/mL; p < 0.05). A positive correlation was found between ATP and GSH (p = 0.014).
The modified cryoprotectant formulation offered superior protection against cryo-induced stress, maintaining higher ATP, GSH, and HSP70 levels post-warming. This formulation holds promise for improving oocyte cryopreservation protocols and conserving the genetic resources of the Kacang goat. Further studies should assess long-term developmental outcomes.
印度尼西亚本土的卡康山羊数量正在减少，这对粮食安全和遗传多样性构成了威胁。成熟和冷冻保存技术是遗传保护的关键策略。然而，冷冻保存过程中的热休克应激会损害卵母细胞的活力。本研究评估了暴露于不同冷冻保护剂下的卡康山羊卵母细胞解冻后的质量。本研究旨在根据成熟率和生物标志物水平（热休克蛋白70 [HSP70]、三磷酸腺苷 [ATP] 和谷胱甘肽 [GSH]），比较改良冷冻保护剂（30% 乙二醇 + 1M 蔗糖）与市售冷冻保护剂在保存解冻后卵母细胞质量方面的效果。
从山羊卵巢中采集卵母细胞并使其成熟22小时。它们被分为三组：对照组（未进行玻璃化）、市售冷冻保护剂组（T1）和改良冷冻保护剂组（T2）。使用酶联免疫吸附测定法评估解冻后的质量，以量化HSP70、ATP和GSH水平。统计分析包括单因素方差分析和Pearson相关性分析（p < 0.05）。
各组的成熟率相当（对照组 [CG]：84.3%，T1：79.8%，T2：77.2%；p > 0.05）。与CG相比，T2组的HSP70水平显著升高（p < 0.05）。与T1组相比，T2组的ATP（52.13 ± 7.7 ng/mL）和GSH（1.27 ± 0.66 ng/mL）水平也显著更高（ATP：25.65 ± 1.63；GSH：0.06 ± 0.01 ng/mL；p < 0.05）。发现ATP与GSH之间存在正相关（p = 0.014）。
改良的冷冻保护剂配方对冷冻诱导的应激提供了更好的保护，解冻后维持了较高的ATP、GSH和HSP70水平。该配方有望改进卵母细胞冷冻保存方案并保护卡康山羊的遗传资源。进一步的研究应评估长期发育结果。
