Evaluation of Different Cryoprotectant Combinations in Vitrification and Slow Freezing for Ovarian Tissue Preservation in Domestic Cats.

IF 1.6 3区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE
Rayane Brandão Ribeiro, Aline Queiroz Rodrigues, Beatriz Alves de Aguiar, Jessyca Karoline Oliveira Silva, Marcus Vinicius Rocha de A Carvalho da Costa, Julliene Larissa Dos Santos Bezerra, Yasmin Barboza Ferreira, Ingrid Gracielle Martins da Silva, Tathyana Benetis Piau, Carolina Madeira Lucci, Sônia Nair Báo, Jair Trapé Goulart, Paula Maria Quaglio Bellozi, Fernanda Paulini
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Abstract

Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol-composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose-proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs.

不同冷冻保护剂组合用于家猫卵巢组织玻璃化和慢速冷冻保存的评价。
在过去的十年中,狩猎活动的增加和栖息地的破坏严重影响了濒临灭绝的狐科动物的数量。认识到这一点,必须实施旨在减轻这种有关保护情况的战略。因此,女性生育能力的保存在这一背景下至关重要,有关这一领域的研究仍然很少。在冷冻保存领域,流行的方法包括慢速冷冻(SF)和玻璃化(V)。本研究旨在评估各种冷冻保护组合对V或SF过程应用于家猫卵巢组织的影响。研究人员解剖了10只健康猫的20个卵巢,将卵巢皮质区域切成8块,每块大小为3mm3。这些片段被随机分配到三个不同的V (V1, V2和V3)或SF (SF1, SF2和SF3)治疗组。每组采用不同浓度的DMSO、EG和海藻糖或蔗糖溶液。评估包括组织学评估、卵泡计数、增殖活性的免疫组织化学分析和超微结构检查。结果表明,V1方案-由10% DMSO, 10% EG和0.1 M海藻糖组成的平衡溶液,然后是20% DMSO, 20% EG和0.1 M海藻糖的V溶液-被证明是最有效的。与其他治疗方法相比,这种组合最好地保存了卵泡形态,减少了变性,支持了卵泡增殖,并保持了良好的超微结构完整性。这些发现为提高家猫的生育能力提供了有价值的基础,在濒危猫科动物保护项目中具有潜在的应用价值。
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来源期刊
Reproduction in Domestic Animals
Reproduction in Domestic Animals 农林科学-奶制品与动物科学
CiteScore
3.00
自引率
5.90%
发文量
238
审稿时长
4-8 weeks
期刊介绍: The journal offers comprehensive information concerning physiology, pathology, and biotechnology of reproduction. Topical results are currently published in original papers, reviews, and short communications with particular attention to investigations on practicable techniques. Carefully selected reports, e. g. on embryo transfer and associated biotechnologies, gene transfer, and spermatology provide a link between basic research and clinical application. The journal applies to breeders, veterinarians, and biologists, and is also of interest in human medicine. Interdisciplinary cooperation is documented in the proceedings of the joint annual meetings. Fields of interest: Animal reproduction and biotechnology with special regard to investigations on applied and clinical research.
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