BIOREN-CEBIOR, Universidad de La Frontera, Temuco, Chile.
Andrologia. 2011 Aug;43(4):233-41. doi: 10.1111/j.1439-0272.2010.01054.x. Epub 2011 Mar 25.
The ability of sucrose to protect spermatozoa against mitochondrial damage, artificial acrosome reaction and DNA fragmentation during ultra-rapid cryopreservation in canine sperm was investigated. Swim-up selected spermatozoa of second-fraction semen were vitrified with different concentrations of sucrose (0.1, 0.25 and 0.4 m) in proportion 1 : 1 v/v with HTF-BSA 1%. From each group, 30-μl suspensions of cells were dropped directly into liquid nitrogen and stored for at least 24 h. Cells were thawed by submerging the spheres in HTF with 1% BSA at 37 °C. The number of progressively motile spermatozoa was significantly higher in the sucrose 0.25 m + HTF-BSA 1% (42.5 ± 2.3%, P < 0.01) than in HTF only (1.66 ± 0.3%). The same combination of sucrose 0.25 m + HTF-BSA 1% (42.7 ± 1.5%) had a stronger cryoprotective effect on the integrity of mitochondrial membrane potential (P < 0.05) and decreased the DNA fragmentation (2.8 ± 0.5%) as compared with HTF only (1.93 ± 0.6% and 5.6 ± 0.6% respectively). With respect to acrosome-reacted spermatozoa, no significant difference was found between the groups investigated (P > 0.05). It is concluded that sucrose, a nonpermeable cryoprotectant, can effectively preserve important physiological parameters such as mitochondrial membrane potential and DNA integrity during ultra-rapid cryopreservation.
研究了蔗糖在犬精子超快速冷冻过程中对精子线粒体损伤、人工顶体反应和 DNA 碎片化的保护作用。用不同浓度的蔗糖（0.1、0.25 和 0.4 m）以 1:1 v/v 的比例与 HTF-BSA 1% 一起对第二部分精液的泳动选择精子进行玻璃化冷冻。从每组中，直接将 30-μl 细胞悬浮液滴入液氮中，并至少储存 24 小时。将球体浸入含 1% BSA 的 HTF 中在 37°C 下解冻。蔗糖 0.25 m+HTF-BSA 1%（42.5±2.3%，P<0.01）组中具有活力的精子数明显高于仅 HTF 组（1.66±0.3%）。蔗糖 0.25 m+HTF-BSA 1%（42.7±1.5%）的组合对线粒体膜电位完整性具有更强的保护作用（P<0.05），并降低 DNA 碎片化（2.8±0.5%），而与仅 HTF 相比（1.93±0.6%和 5.6±0.6%）。就顶体反应的精子而言，各组之间没有发现显著差异（P>0.05）。结论是，蔗糖作为一种不可渗透的冷冻保护剂，在超快速冷冻过程中可以有效地保存重要的生理参数，如线粒体膜电位和 DNA 完整性。
