Carvalho Julyne Vivian Guimarães de, Soares Airton R B, Leão Danuza L, Reis Adriana N, Santos Regiane R, Rodrigues Ana P R, Domingues Sheyla F S
Laboratory of Wild Animal Biotechnology and Medicine, Faculty of Veterinary Medicine, Federal University of Pará, Castanhal 68740-970, Brazil.
Postgraduate Program in Animal Health and Production in the Amazon, Federal Rural University of the Amazon, Belém 66077-830, Brazil.
Animals (Basel). 2023 Aug 31;13(17):2768. doi: 10.3390/ani13172768.
Vitrification is essential for successful tissue cryopreservation and biobanking in wild cats. This study aimed to compare different methods of vitrification (Ovarian Tissue Cryosystem-OTC, Straws-STW, and Solid Surface vitrification-SSV) for testicular fragment vitrification in tom cats. Testicular fragments were recovered from five adult tom cats and subjected to equilibrium vitrification using different cryovials and methods under the same conditions of vitrification solutions and cryoprotectants. The efficiencies of the methods were evaluated using histological analysis of spermatogonia and Sertoli cell nuclei, seminiferous tubular basement membrane detachment, and the gonadal epithelium shrinkage score scale. Cell viability was assessed using Hoechst PI and Terminal deoxynucleotidyl transferase nick end labeling (TUNEL) assay. The results showed that OTC is an effective vitrification method for maintaining the distinction between spermatogonia and Sertoli cells. OTC was similar to the control for basal membrane detachment parameters ( = 0.05). Epithelial shrinkage was low in the SSV group, which showed the highest percentage of viable cells among the vitrified groups ( = 0.0023). The OTC and SSV vitrification methods were statistically similar in terms of the percentage of TUNEL-positive cells ( = 0.05). Therefore, OTC and SSV provide favorable conditions for maintaining viable cat testicular tissue cells after vitrification.
玻璃化对于野猫组织的成功冷冻保存和生物样本库建设至关重要。本研究旨在比较不同的玻璃化方法（卵巢组织冷冻系统-OTC、细管-STW和固体表面玻璃化-SSV）用于雄猫睾丸片段的玻璃化。从五只成年雄猫获取睾丸片段，并在相同的玻璃化溶液和冷冻保护剂条件下，使用不同的冻存管和方法进行平衡玻璃化。通过对精原细胞和支持细胞核的组织学分析、生精小管基底膜脱离情况以及性腺上皮收缩评分量表来评估这些方法的效率。使用Hoechst PI和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）检测来评估细胞活力。结果表明，OTC是一种有效的玻璃化方法，可维持精原细胞和支持细胞之间的区别。OTC在基底膜脱离参数方面与对照组相似（P = 0.05）。SSV组上皮收缩程度低，在玻璃化组中显示出最高的活细胞百分比（P = 0.0023）。在TUNEL阳性细胞百分比方面，OTC和SSV玻璃化方法在统计学上相似（P = 0.05）。因此，OTC和SSV为玻璃化后维持猫睾丸组织活细胞提供了有利条件。
