IMPACT OF A COMMERICALLY AVAILABLE SPERM SEPERATION DEVICE (ZYMŌT MULTI) ON SPERM DNA QUALITY COMPARED TO DENSITY GRADIENT SEPERATION WITH SWIM UP

Abstract

Isolation of high quality sperm is a pre-requisite for IVF/ICSI to ensure proper fertilization, embryo development and to obtain successful clinical outcomes. The quality of sperm not only entails sufficient motility and morphology, but also involves isolating sperm with high DNA integrity. Various sperm isolation methods exist and some may be superior to others with regard to obtaining sperm with intact DNA. This study compared DNA integrity of sperm isolation using a novel sperm separation device that uses sperm motility and a separation membrane (ZYMōT Multi) or the traditional density gradient separation (DGS) followed by swim-up approach. Prospective trial Semen samples were obtained from 30 men. All samples were split and processed using both the ZYMōT MULTI device as well as DGS + swim-up. Briefly, the ZYMōT preparation entailed adding raw semen to the device inlet port and obtaining isolated sperm from the outlet. DGS + swim-up entailed layer raw semen over the top of a 40%/80% gradient (Puresperm) and spinning the sample at 300xg for 20 minutes followed by two 8 minute washes (Quinn’s sperm wash) at 300xg. The pellet was then overlaid by GIVF medium and motile sperm isolated from the top fraction ∼1hr later. DNA fragmentation (DFI) was assessed on all isolated samples and also to the raw semen using the TUNEL assay. Data were analyzed using ANOVA and Tukey analysis, p<0.05. The DFI of the raw semen was assessed. Use of both sperm separation techniques examined in this study resulted in isolation of sperm with significantly improved sperm DNA integrity compared to sperm from raw semen. There was no statistically difference in in resulting sperm DFI levels between use of ZYMōT MULTI and DGS + swim up.Tabled 1Raw SemenDGS +Swim-upZYMōT MULTIAvg Sperm DFI19.2 ±2.4a7.2±0.9b6.7±0.9bdifferent superscripts represent statistically significant differences between treatments, p<0.05 Open table in a new tab different superscripts represent statistically significant differences between treatments, p<0.05 Different sperm separation techniques can improve sperm DNA integrity compared to sperm found in raw semen. Use the commercial sperm separation device provided no increase in improvement in resulting sperm DNA fragmentation compared to careful sampling of sperm prepared via proper DGS followed by swim-up. Future studies will examine impact of the novel sperm separation device on resulting fertilization, embryo development/aneuploidy and clinical outcomes.