Rahman M A, Park S H, Yu I J
Laboratory of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-safety Research Institute, Chonbuk National University, Iksan, Republic of Korea.
Cryo Letters. 2017 Jan/Feb;38(1):51-57.
The present study investigated the effect of monosaccharides in a glycerol-free tris (GFT) extender on reactive oxygen species (ROS) levels and apoptosis in cryopreserved dog spermatozoa.
The sperm pellets were resuspended (5 × 10 mL) with GFT containing 172 mM glucose (G), 86 mM glucose + 86 mM fructose (GF), 86 mM glucose + 86 mM galactose (Gg), 172 mM fructose (F), 172 mM galactose (g) or 86 mM fructose + 86 mM galactose (Fg). The sperm (500 µL) were loaded in 0.5 mL straws and cooled for 50 min at 4 degree C. The straws were then frozen 7 cm from the surface of liquid nitrogen (LN2) for 20 min and were finally plunged into LN2. After freezing-thawing, the sperm motility and viability were evaluated. The ROS level (HO) and apoptosis index were assessed by using flow cytometry.
GFT supplemented with GF resulted in significantly higher (P < 0.05) progressive sperm motility and viability followed by only G, which had greater ROS reducing capacity. However, sperm cell apoptosis had no significant differences among all the experimental groups.
Cryopreservation of dog sperm in GFT containing GF yields more motile and viable sperm followed by only G, which has greater ROS reducing efficiency.
本研究调查了无甘油三羟甲基氨基甲烷（GFT）稀释液中的单糖对冷冻保存的犬精子活性氧（ROS）水平和凋亡的影响。
将精子沉淀重悬（5×10 mL）于含有172 mM葡萄糖（G）、86 mM葡萄糖 + 86 mM果糖（GF）、86 mM葡萄糖 + 86 mM半乳糖（Gg）、172 mM果糖（F）、172 mM半乳糖（g）或86 mM果糖 + 86 mM半乳糖（Fg）的GFT中。将500 μL精子装入0.5 mL细管中，在4℃下冷却50分钟。然后将细管在距液氮（LN2）表面7 cm处冷冻20分钟，最后投入LN2中。冻融后，评估精子活力和存活率。使用流式细胞术评估ROS水平（HO）和凋亡指数。
添加GF的GFT导致显著更高（P < 0.05）的精子渐进性活力和存活率，其次是仅添加G的情况，G具有更大的ROS还原能力。然而，所有实验组之间的精子细胞凋亡没有显著差异。
在含有GF的GFT中冷冻保存犬精子可产生更多有活力和存活的精子，其次是仅添加G的情况，G具有更高的ROS还原效率。
