从冷冻解冻的水牛精液中分离活动精子：上浮法与过滤法

Separation of motile spermatozoa from frozen-thawed buffalo semen: swim-up vs filtration procedures.

作者信息

Mustafa G, Anzar M, Arslan M

机构信息

Department of Biological Sciences, Quaid-i-Azam University, Pakistan.

出版信息

Theriogenology. 1998 Jul 15;50(2):205-11. doi: 10.1016/s0093-691x(98)00127-7.

DOI:10.1016/s0093-691x(98)00127-7

PMID:10734487

Abstract

Three experiments were conducted to maximize the recovery rate of motile spermatozoa from frozen-thawed buffalo semen. In Experiment 1, the swim-up of motile spermatozoa was performed in the presence or absence of HEPES in TALP medium and CO2 in the environment. The recovery rate of motile spermatozoa in TALP medium (control), TALP + HEPES + CO2, TALP + HEPES and TALP + CO2 was 15, 18, 12 and 10%, respectively (P > 0.05), with sperm motility at 87, 89, 90 and 90%, respectively (P > 0.05). In Experiment 2, the pH of TALP medium was adjusted to 7.0, 7.5, 8.0, 8.5 and 9.0, and swim-up procedure was performed in the presence of HEPES and CO2. The recovery rate of motile spermatozoa at different pH was 14, 20, 24, 27 and 16%, respectively (P < 0.05). Motility of separated spermatozoa was 88, 91, 90, 89 and 90%, respectively (P > 0.05). In Experiment 3, the efficiency of ion-exchange filtration and Swim-up procedure in separating motile spermatozoa from frozen-thawed buffalo semen was compared. The recovery rate of motile spermatozoa was 95% in filtration procedure and 33% in swim-up procedure (P < 0.005). In all experiments, normal acrosomes did not vary due to treatments (P > 0.05). In conclusion, HEPES and CO2 had no significant effect on swim-up of buffalo spermatozoa. The pH 8.5 of TALP improved the recovery rate of motile spermatozoa in swim-up procedure. The ion-exchange filtration was found superior to swim-up procedure in harvesting maximum number of motile spermatozoa from frozen-thawed buffalo semen (95 vs 33%; P < 0.001).

摘要

进行了三项实验，以最大限度地提高从冻融水牛精液中回收活动精子的比率。在实验1中，在TALP培养基中添加或不添加HEPES以及环境中添加或不添加CO₂的情况下，对活动精子进行上浮分离。TALP培养基（对照）、TALP + HEPES + CO₂、TALP + HEPES和TALP + CO₂中活动精子的回收率分别为15%、18%、12%和10%（P>0.05），精子活力分别为87%、89%、90%和90%（P>0.05）。在实验2中，将TALP培养基的pH值调整为7.0、7.5、8.0、8.5和9.0，并在添加HEPES和CO₂的情况下进行上浮分离操作。不同pH值下活动精子的回收率分别为14%、20%、24%、27%和16%（P<0.05）。分离出的精子活力分别为88%、91%、90%、89%和90%（P>0.05）。在实验3中，比较了离子交换过滤和上浮分离操作从冻融水牛精液中分离活动精子的效率。过滤操作中活动精子的回收率为95%，上浮分离操作中为33%（P<0.005）。在所有实验中，正常顶体不受处理影响（P>0.05）。总之，HEPES和CO₂对水牛精子的上浮分离没有显著影响。TALP培养基pH值为8.5时，可提高上浮分离操作中活动精子的回收率。发现在从冻融水牛精液中获取最大数量的活动精子方面，离子交换过滤优于上浮分离操作（95%对33%；P<0.001）。