Palazzese Luca, Turri Federica, Anzalone Debora Agata, Saragusty Joseph, Bonnet Jacques, Colotte Marthe, Tuffet Sophie, Pizzi Flavia, Luciani Alessia, Matsukawa Kazutsugu, Czernik Marta, Loi Pasqualino
Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, Warsaw, Poland.
Institute of Agricultural Biology and Biotechnology (IBBA), National Research Council (CNR), Lodi, Italy.
Front Vet Sci. 2023 Nov 30;10:1270266. doi: 10.3389/fvets.2023.1270266. eCollection 2023.
Freeze-drying techniques give alternative preservation mammalian spermatozoa without liquid nitrogen. However, most of the work has been conducted in the laboratory mouse, while little information has been gathered on large animals that could also benefit from this kind of storage.
This work adapted a technique known as vacuum-drying encapsulation (VDE), originally developed for nucleic acid conservation in anhydrous state, to ram spermatozoa, and compared it to canonical lyophilization (FD), testing long-term storage at room temperature (RT) and 4°C.
The results demonstrated better structural stability, namely lipid composition and DNA integrity, in VDE spermatozoa than FD ones, with outcomes at RT storage comparable to 4°C. Likewise, in VDE the embryonic development was higher than in FD samples (12.8% vs. 8.7%, < 0.001, respectively). Our findings indicated that in large mammals, it is important to consider dehydration-related changes in sperm polyunsaturated fatty acids coupled with DNA alterations, given their crucial role in embryonic development.
冷冻干燥技术为哺乳动物精子提供了无需液氮的替代保存方法。然而，大部分研究工作是在实验室小鼠身上进行的，而对于同样能从这种保存方式中受益的大型动物，所收集的信息却很少。
本研究采用了一种名为真空干燥封装（VDE）的技术，该技术最初是为无水状态下的核酸保存而开发的，将其应用于公羊精子，并与传统冻干法（FD）进行比较，测试在室温（RT）和4°C下的长期保存效果。
结果表明，VDE处理的精子在结构稳定性方面表现更好，即脂质组成和DNA完整性方面优于FD处理的精子，室温保存的效果与4°C保存相当。同样，VDE处理的胚胎发育率高于FD处理的样本（分别为12.8%和8.7%，<0.001）。我们的研究结果表明，在大型哺乳动物中，鉴于精子多不饱和脂肪酸的脱水相关变化以及DNA改变在胚胎发育中的关键作用，考虑这些因素非常重要。
