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使用不同稀释液和冷冻保护剂对长颈鹿附睾精子进行冷冻保存

Cryopreservation of Giraffe Epidydimal Spermatozoa Using Different Extenders and Cryoprotectants.

作者信息

Hermes Robert, Lecu Alexis, Potier Romain, Goeritz Frank, Rickard Jessica P, Bohner Julia, Wedlarski Rudy, Hruby Jiri, Hildebrandt Thomas B

机构信息

Department Reproduction Management, Leibniz Institute for Zoo and Wildlife Research, D-10315 Berlin, Germany.

Parc Zoologique de Paris, MNHN, F-75012 Paris, France.

出版信息

Animals (Basel). 2022 Mar 29;12(7):857. doi: 10.3390/ani12070857.

Abstract

Giraffe numbers have plummeted over the last 30 years by 30-40%. Thus, their conservation status has been raised from least concern to vulnerable. Efforts to manage in situ and ex situ populations are increasing. Assisted reproduction techniques (ART) such as sperm cryopreservation could help preserve the genetic diversity of giraffe subspecies and, when used for artificial inseminations, enhance genetic exchange between isolated populations. However, to date, the post-thaw motility of recovered sperm has been low and inconsistent. In this study, epididymal sperm collected from the testes of giraffes ( = 7) was frozen in three different extenders, namely, BotuCrio, Steridyl, and test egg yolk (TEY), each supplemented with one of two different cryoprotectants (5% glycerol or a mix of 1% glycerol and 4% methylformamide) and frozen over liquid nitrogen vapor. Across all three extenders, sperm showed significantly better post-thaw results when frozen with a mix of glycerol and methylformamide compared with glycerol alone. Sperm frozen with TEY and a mix of glycerol and methylformamide achieved superior post-thaw total and progressive sperm motility of 57 ± 3% and 45 ± 3%, respectively. These results show the benefit of using alternative cryoprotectants for freezing giraffe spermatozoa and could aid in the application of ARTs for giraffe subspecies or the closely related endangered Okapi.

摘要

在过去30年里,长颈鹿数量锐减了30%至40%。因此,它们的保护等级已从“最无危”提升为“易危”。针对长颈鹿种群的原地和迁地管理工作正在增加。诸如精子冷冻保存等辅助生殖技术(ART)有助于保护长颈鹿亚种的遗传多样性,并且用于人工授精时,能促进隔离种群间的基因交流。然而,迄今为止,复苏精子的解冻后活力一直很低且不稳定。在本研究中,从7头长颈鹿睾丸采集的附睾精子被冷冻保存在三种不同的稀释液中,即BotuCrio、Steridyl和试验蛋黄(TEY),每种稀释液均添加两种不同冷冻保护剂之一(5%甘油或1%甘油与4%甲酰胺的混合物),并在液氮蒸汽上冷冻。在所有三种稀释液中,与仅使用甘油相比,当使用甘油和甲酰胺混合物冷冻时,精子解冻后的结果显著更好。用TEY以及甘油和甲酰胺混合物冷冻的精子解冻后总活力和渐进性活力分别达到了57±3%和45±3%的优异水平。这些结果表明了使用替代冷冻保护剂冷冻长颈鹿精子的益处,并有助于将辅助生殖技术应用于长颈鹿亚种或与之密切相关的濒危霍加狓。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f5/8997136/bcac0fcdc912/animals-12-00857-g001.jpg

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