Matás C, Decuadro G, Martínez-Miró S, Gadea J
Department of Physiology, School of Veterinary, University of Murcia, Spain.
Theriogenology. 2007 Mar 15;67(5):1087-91. doi: 10.1016/j.theriogenology.2006.11.010. Epub 2006 Dec 18.
The purpose of this investigation was to evaluate the use of an iodixanol cushion during centrifugation on sperm recovery and yield after centrifugation (sperm recovery, sperm motility, viability, membrane lipid disorder, acrosome reaction and ROS generation); and to investigate how this procedure affects sperm function after freezing-thawing (sperm motility, membrane lipid disorder, acrosomal status and homologous in vitro penetration test). The sperm-rich fractions from fertile boars were centrifuged under two centrifugation régimes: 800xg for 10min (standard method) and 1000xg for 20min with an iodixanol (60% w/v) cushion at the bottom of the centrifuge tubes (Cushion method). The highest recovery was achieved using the cushion method (sperm loss for cushion method was 0.50%+/-0.18 versus 2.97%+/-0.43 for standard method, P<0.01) and sperm quality was not significantly affected by the centrifugation régime. The motion parameters (% progressive motility, % motility, VCL, VSL, VAP, ALH, BCF, P<0.05) of frozen-thawed samples showed higher values using the standard method. However, a higher number of viable spermatozoa with lower lipid disorders were found in spermatozoa processed with the cushion method. The in vitro penetration assay showed that the individual boar influenced the parameters studied but there were no differences between the two centrifugation régimes used. Our results support the hypothesis that the proportion of sperm loss in frozen-thawed semen was significantly influenced by the centrifugation régime. Therefore, the iodixanol cushion method is a suitable tool for cryopreservation of boar semen in order to reduce sperm loss without affecting sperm quality.
本研究的目的是评估在离心过程中使用碘克沙醇垫层对精子回收率以及离心后产量（精子回收率、精子活力、存活率、膜脂紊乱、顶体反应和活性氧生成）的影响；并研究该方法对冻融后精子功能（精子活力、膜脂紊乱、顶体状态和同源体外穿透试验）的影响。将来自可育公猪的富含精子的部分在两种离心条件下进行离心：800xg离心10分钟（标准方法）和1000xg离心20分钟，在离心管底部使用碘克沙醇（60% w/v）垫层（垫层法）。使用垫层法获得了最高的回收率（垫层法的精子损失率为0.50%±0.18，而标准方法为2.97%±0.43，P<0.01），并且离心条件对精子质量没有显著影响。冻融样品的运动参数（% 前向运动率、% 运动率、曲线速度、直线速度、平均路径速度、平均侧摆幅度、鞭打频率，P<0.05）在使用标准方法时显示出更高的值。然而，在用垫层法处理的精子中发现了更多具有较低脂质紊乱的活精子。体外穿透试验表明，个体公猪会影响所研究的参数，但所使用的两种离心条件之间没有差异。我们的结果支持以下假设，即冻融精液中精子损失的比例受离心条件的显著影响。因此，碘克沙醇垫层法是一种适用于公猪精液冷冻保存的工具，以减少精子损失而不影响精子质量。
