Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, Cuenca, Ecuador.
Laboratorio de Sanidad Animal y Zoonosis, Departamento de Ciencias Biológicas y Agropecuarias, Universidad Técnica Particular de Loja, Loja, Ecuador.
Biopreserv Biobank. 2024 Apr;22(2):157-165. doi: 10.1089/bio.2022.0214. Epub 2023 Aug 17.
This study aimed to assess the suitability of egg yolk (EY) supplementation to a tris-citric acid-based extender on cryosurvival of guinea pig () epididymal spermatozoa. Two synthetic-based extenders, tris-citric acid-glucose plus 20% EY (TCG-EY) and tris-citric acid-fructose (TCF) both with 5% glycerol, were compared. Thirty-two epididymides were recovered from 16 adult guinea pig males by gonadectomy, and then the sperm samples were retrieved by retrograde flushing using TCG-EY and TCF extenders for left or right epididymis, respectively. TCG-EY and TCF sperm samples were frozen in static liquid nitrogen vapors through a two-step cooling procedure. Before freezing, the percentage of progressive sperm motility and sperm with intact plasma and acrosome membranes from TCG-EY sperm samples were higher ( < 0.05) than those diluted with TCF. Post-thaw sperm kinematic variables and membrane integrity were drastically reduced ( < 0.001) compared with prefreezing samples, regardless of extender type. The post-thaw plasmatic and acrosome membrane integrity from TCG-EY sperm samples was higher ( < 0.05) than those from TCF samples. Except for the length, the morphometric head dimensions of sperm diluted with TCG-EY or TCF did not vary ( > 0.05) after the freezing-thawing process compared with the prefreezing samples. In conclusion, despite greater cell cryoinjury with both extenders, the EY supplementation exerted greater cell membrane protection before and after the freezing-thawing process. This research shows an in-depth analysis of guinea pig sperm cryopreservation; however, more studies are recommended.
本研究旨在评估卵黄(EY)补充到基于三柠檬酸的稀释液中对豚鼠附睾精子冷冻保存的适用性。比较了两种基于合成物的稀释液:三柠檬酸-葡萄糖加 20%EY(TCG-EY)和三柠檬酸-果糖(TCF),两者均含有 5%甘油。从 16 只成年雄性豚鼠的性腺切除术回收 32 个附睾,然后通过逆行冲洗用 TCG-EY 和 TCF 稀释液分别回收左侧或右侧附睾的精子样本。TCG-EY 和 TCF 精子样本通过两步冷却程序在静态液氮蒸气中冷冻。在冷冻前,TCG-EY 精子样本的渐进精子运动百分比和具有完整质膜和顶体膜的精子比例高于用 TCF 稀释的精子样本(<0.05)。与冷冻前样本相比,无论稀释液类型如何,解冻后精子运动学变量和膜完整性都急剧降低(<0.001)。与 TCF 样本相比,TCG-EY 精子样本解冻后的质膜和顶体膜完整性更高(<0.05)。除了长度外,用 TCG-EY 或 TCF 稀释的精子的形态学头部尺寸在冷冻-解冻过程后与冷冻前样本相比没有变化(>0.05)。总之,尽管两种稀释液都对细胞造成了更大的冷冻损伤,但 EY 补充在冷冻-解冻前后对细胞膜起到了更大的保护作用。这项研究对豚鼠精子冷冻保存进行了深入分析,但建议进行更多研究。