FINE-TUNING SPEED AND TIME OF CENTRIFUGATION FOR SPERM WASH PROCESS IN MALE FACTOR

Abstract

The objective of this study is to determine whether speed and/or time of centrifugation for sperm wash process effects on the motility of harvested sperm and total motile sperm count. DESIGN: Among donated frozen sperm for quality control purpose, male factor sperm was used in this study. According to WHO 5th edition, sperm wash for diluted semen sample should be centrifuged at 300-500g for 5-10 minutes. Six experimental condition was set as Ex1: 500g for 10 min, Ex2: 400g for 10 min, Ex3: 300g for 10 min, Ex4: 500g for 5 min, Ex5: 400g for 5 min, Ex6: 300g for 5 min. Each experiments were repeatedly performed five times using same centrifuge (Eppendorf 5702). Donated frozen sperm were thawed for 20 min at 37°C. Equal volume of pre-warmed sperm wash was added into thawed sperm and six aliquots were prepared for experiment. Average sperm motility was 17% (range: 9% to 30%) and average progressive motility was 12% (range: 6% to 20%). We analyzed the difference of motility of harvested sperm after single centrifugation and Ex1 was used as control group. The ANOVA and t-test were used. A p value <0.05 was considered significant. After six different condition of centrifugation, manual sperm count for motility was performed by one andrologist to reduce personal bias. Comparing to Ex1, sperm motility in Ex3 and Ex6 was significantly higher than that of other groups (p=0.028 and p=0.013, respectively). Progressive motility in Ex3 and Ex6 was also significantly higher than other groups (p=0.012 and p=0.0003). There is no significant difference of sperm motility between Ex3 and Ex6. Between 300g and 500g for 5 min to 10 min range, 300g for 5 min was sufficient for sperm wash with keeping good sperm motility. After six different condition of centrifugation, manual sperm count for motility was performed by one andrologist to reduce personal bias. Comparing to Ex1, sperm motility in Ex3 and Ex6 was significantly higher than that of other groups (p=0.028 and p=0.013, respectively). Progressive motility in Ex3 and Ex6 was also significantly higher than other groups (p=0.012 and p=0.0003). There is no significant difference of sperm motility between Ex3 and Ex6. Between 300g and 500g for 5 min to 10 min range, 300g for 5 min was sufficient for sperm wash with keeping good sperm motility.