Vizuete G, Jiménez E, Agüera E I, Pérez-Marín C C
Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Cordoba, Spain.
Reprod Domest Anim. 2014 Feb;49(1):e5-8. doi: 10.1111/rda.12253. Epub 2013 Nov 21.
The objective of the present study was to investigate the influence of different sucrose-based extenders on the motility, morphology, viability and acrosomal integrity of epididymal cat spermatozoa cryopreserved by ultra-rapid freezing method. Nine cats were castrated, and collected semen was diluted 1 : 1 with Dulbecco`s phosphate-buffered saline-BSA1%-based extender supplemented with different sucrose concentrations (0, 0.25, 0.4 and 0.6 m). After ultra-rapid freezing, samples were thawed and sperm motility, morphology, viability and acrosome status were assessed. At thawing, the number of progressively motile (p < 0.01) and morphologically normal (p < 0.01) sperm was higher in the sucrose-supplemented groups than in the sucrose-free group. Viability of spermatozoa cryopreserved without sucrose was significantly reduced. In extender supplemented with 0.4 m sucrose, spermatozoa viability showed higher values (57.0 ± 4.7; p < 0.01). No significant differences were detected among groups for sperm acrosome integrity. Results support that cat sperm survive after ultra-rapid freezing using sucrose as a cryoprotectant, and the best results were achieved when 0.4 m of sucrose was used. This is the first report on sperm ultra-rapid freezing of cat sperm and further studies on extenders, sperm management or cryovials should be carried out to improve sperm cryosurvival.
本研究的目的是调查不同蔗糖基稀释液对采用超快速冷冻法冷冻保存的附睾猫精子活力、形态、存活率和顶体完整性的影响。对9只猫实施去势手术，采集的精液用添加不同蔗糖浓度（0、0.25、0.4和0.6 m）的基于杜氏磷酸盐缓冲盐水-牛血清白蛋白1%的稀释液按1:1进行稀释。超快速冷冻后，将样品解冻并评估精子活力、形态、存活率和顶体状态。解冻时，添加蔗糖的组中进行性运动（p < 0.01）和形态正常（p < 0.01）的精子数量高于无蔗糖组。未添加蔗糖冷冻保存的精子存活率显著降低。在添加0.4 m蔗糖的稀释液中，精子存活率显示出较高值（57.0 ± 4.7；p < 0.01）。各组间精子顶体完整性未检测到显著差异。结果支持猫精子在以蔗糖作为冷冻保护剂进行超快速冷冻后能够存活，并且使用0.4 m蔗糖时取得了最佳结果。这是关于猫精子超快速冷冻的首篇报道，应进一步开展关于稀释液、精子处理或冻存管的研究以提高精子冷冻存活率。
