Abstract Long-term storage of spermatogonial stem cells (SSCs) represents the next step for advancing aquaculture research by providing valuable genetic resources for genetic enhancement programs and xenogenesis technologies. Hybrid catfish, the cross between channel catfish, Ictalurus punctatus ♀ and blue catfish, I. furcatus ♂, are in high demand by the US aquaculture industry, but production can be limited by the availability of blue catfish gametes. Therefore, xenogeneic stem cell transplantation is a powerful method for generating blue catfish gametes within faster growing and maturing channel catfish hosts. These technologies could be facilitated by having cryopreserved SSCs from testicular tissue on-hand in gene banks. Cryopreservation protocols for blue catfish testicular tissues and for other fish species are still being optimized, and currently, data is lacking on the effects antioxidants and antifreeze proteins (AFPs) have on cellular post-thaw viability of SSCs. The objective of this study was to analyze the individual and combined effects of antioxidants (catalase, hypotaurine, and ascorbic acid) and AFPs (I and III) on post-thaw type A spermatogonia cell production and viability from testicular tissue. In this study, there were no improvements with individual antioxidant or AFP treatments. However, when the antioxidants hypotaurine and catalase were tested in combination with different AFPs, three treatments had higher type A spermatogonia cell production than the control, including hypotaurine 7 mM + AFPI 1 μg/mL, hypotaurine 3.5 mM + AFPI 0.1 μg/mL, and hypotaurine 7 mM + AFPIII 0.1 μg/mL. These treatments along with hypotaurine 7 mM + AFPI 0.1 μg/mL and catalase 100 IU + AFPI 0.1 μg/mL had higher viability than the control. From these results, we recommend adding these antioxidant-AFP combinations to future cryopreservation protocols. Overall, this is the first study showing that combining specific antioxidants and AFPs can improve spermatogonia cryopreservation for a fish species.

中文翻译：

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抗氧化剂和抗冻蛋白对蓝鲶（Ictalurus furcatus）精原细胞冷冻保存的影响

摘要 精原干细胞 (SSC) 的长期储存代表了通过为遗传增强计划和异种技术提供宝贵的遗传资源来推进水产养殖研究的下一步。美国水产养殖业对杂交鲶鱼（沟鲶 Ictalurus punctatus ♀ 和蓝鲶鱼 I. furcatus ♂ 的杂交）的需求量很大，但产量可能受到蓝鲶配子的可用性的限制。因此，异种干细胞移植是在快速生长和成熟的沟鲶宿主内生成蓝鲶配子的有效方法。这些技术可以通过在基因库中从睾丸组织中冷冻保存 SSC 来促进。蓝鲶睾丸组织和其他鱼类的冷冻保存方案仍在优化中，目前，缺乏关于抗氧化剂和抗冻蛋白 (AFP) 对 SSC 细胞解冻后活力影响的数据。本研究的目的是分析抗氧化剂（过氧化氢酶、亚牛磺酸和抗坏血酸）和 AFP（I 和 III）对睾丸组织解冻后 A 型精原细胞的产生和活力的单独和联合作用。在这项研究中，单独的抗氧化剂或 AFP 治疗没有改善。然而，当抗氧化剂亚牛磺酸和过氧化氢酶与不同的 AFP 联合测试时，三种处理的 A 型精原细胞产量高于对照，包括亚牛磺酸 7 mM + AFPI 1 μg/mL、亚牛磺酸 3.5 mM + AFPI 0.1 μg/mL，和亚牛磺酸 7 mM + AFPIII 0.1 μg/mL。这些治疗与亚牛磺酸 7 mM + AFPI 0 一起使用。1 μg/mL 和过氧化氢酶 100 IU + AFPI 0.1 μg/mL 比对照具有更高的活力。根据这些结果，我们建议将这些抗氧化剂-AFP 组合添加到未来的冷冻保存方案中。总的来说，这是第一项表明结合特定抗氧化剂和 AFP 可以改善鱼类精原细胞冷冻保存的研究。