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使用蔗糖作为冷冻保护剂,通过不同方法(珠粒法和迷你细管法)对猪精子进行玻璃化冷冻后,成功恢复了其活力和生存能力。

Successful recovery of motile and viable boar sperm after vitrification with different methods (pearls and mini straws) using sucrose as a cryoprotectant.

作者信息

Serrano Albal María, Aquilina Marie Claire, Zak Louisa J, Ellis Peter J, Griffin Darren K

机构信息

School of Biosciences, University of Kent, Canterbury, United Kingdom.

School of Biosciences, University of Kent, Canterbury, United Kingdom.

出版信息

Cryobiology. 2023 Dec;113:104583. doi: 10.1016/j.cryobiol.2023.104583. Epub 2023 Aug 31.

Abstract

Vitrification of sperm by direct contact with liquid nitrogen is increasing in popularity as an alternative to conventional (slow) freezing. Although slow freezing is very challenging in boar sperm cryopreservation, this is currently the standard method used. We compared vitrification in "pearls" and in "mini straws" using the in vitro fertilization media Porcine Gamete Media with 0.3 M sucrose with the standard (slow) method used to preserve boar sperm. Both vitrification methods reduced the viability of the sperm sample more than slow freezing (42.2 ± 4.3% total motility and 71.4 ± 2.3% alive), however, both protocols allowed for the successful recovery of the sperm samples. By comparing two different methods of vitrification and two different methods of post-thaw preparation we were able to determine the optimal vitrification-thaw protocol for boar sperm. When comparing pearls and mini-straws, the smaller liquid volume associated with pearls had a positive effect on the survivability of the samples, reducing sperm DNA damage (1.2 ± 0.2% vs. 5.1 ± 0.1.7%) and preserving motility (26.15 ± 2.8% vs 9.39 ± 0.9%) after thawing. In conclusion, the pearl method was the most suitable of the vitrification techniques for use with boar sperm.

摘要

与液氮直接接触对精子进行玻璃化冷冻作为传统(慢速)冷冻的替代方法正越来越受欢迎。尽管慢速冷冻在公猪精子冷冻保存中极具挑战性,但目前仍是使用的标准方法。我们使用含0.3M蔗糖的体外受精培养基猪配子培养基,将“珍珠”和“迷你细管”中的玻璃化冷冻方法与用于保存公猪精子的标准(慢速)方法进行了比较。两种玻璃化冷冻方法比慢速冷冻使精子样本的活力降低得更多(总活力为42.2±4.3%,活率为71.4±2.3%),然而,两种方案都能成功回收精子样本。通过比较两种不同的玻璃化冷冻方法和两种不同的解冻后处理方法,我们能够确定公猪精子的最佳玻璃化冷冻-解冻方案。在比较“珍珠”和“迷你细管”时,与“珍珠”相关的较小液体体积对样本的存活率有积极影响,解冻后减少了精子DNA损伤(分别为1.2±0.2%和5.1±0.17%)并保持了活力(分别为26.15±2.8%和9.39±0.9%)。总之,“珍珠”法是最适合用于公猪精子的玻璃化冷冻技术。

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