DOES ASSISTED LIQUEFACTION INSTEAD OF INCUBATION AFFECT SPERM MOTILITY AND MOTILE SPERM RECOVERY IN THE SPERM WASH PROCESS?

Abstract

The primary purpose of sperm preparation may be to separate motile sperm from seminal plasma as soon as possible and prepare an optimal sample for ART, such as in vitro fertilization (IVF) and intrauterine insemination (IUI). The objective of this study is to evaluate whether an assisted liquefaction technique can replace the traditional method and whether sperm motility and motile sperm recovery is affected. Twenty samples were analyzed using both traditional and assisted liquefaction methods. The assisted liquefaction technique used consisted of adding pre-warmed sperm wash media at 1 to 2x the semen volume to the sample before sample processing. Then liquefaction and viscosity were confirmed by dropping specimen. Half of the sample was liquefied for 15-30 minutes at 37°C outlined by WHO 5th edition. Each sample was then overlaid onto 2 ml of 90% density gradient with no more than 1.5 ml of sample per tube. The sample was centrifuged at 400g for 15 minutes. Supernatant was removed and samples from multiple tubes were combined if needed. After additional wash, a 5 ul aliquot from both group was counted using a pre-warmed MicroCell as a post-wash. The difference between the assisted and traditional liquefaction was assessed for the following parameters: total motile sperm, total motility, and progressive motility by t-test. A p value of < 0.05 was considered significant. The assisted liquefaction resulted in an average increase of 3% for both the total motility and progressive motility and recovered in an average increase of 1.69 million motile sperm although there was not significantly different. Furthermore, sperm were observed to move at a higher velocity in samples processed with the assisted technique than with the traditional by experienced andrologists. Samples processed with the traditional technique were observed to contain more non-motile sperm after processing than those processed with the assisted technique The use of an assisted technique instead of the traditional technique can be used without negatively affecting motility and recovery of motile sperm for ART. An assisted technique can be used to decrease the processing time while continuing to provide a high quality sample. Additionally, an assisted technique may provide a more efficient sample preparation as the seminal plasma is fully diluted resulting in less non-motile sperm and greater progressive motility.