Comparative analysis of density gradient centrifugation and magnetic-activated cell sorting for the enrichment of X- and Y-chromosome-linked spermatozoa and their influence on pregnancy rates

Abstract

The desire to have offspring of a specific sex has a long history, becoming especially prevalent with the advent of assisted reproductive technologies (ARTs). This study aimed to compare the effectiveness of two sperm X and Y Chromosome-Bearing Selection techniques, density gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS), in enriching Y-bearing sperm for intracytoplasmic sperm injection (ICSI), analyzed by the fluorescence in-situ hybridization (FISH) technique. Sixty patients undergoing ICSI were enrolled. Their spermatozoa were prepared using either DGC or MACS. After preparation with Ham’s F10 medium, X- and Y-chromosome-bearing spermatozoa were identified and separated using three methods: wash (control), DGC, and MACS. The FISH technique was employed to quantify the proportion of Y-chromosome-bearing sperm. X- and Y-chromosome-bearing spermatozoa were identified by the presence of green or orange fluorescent signals, respectively. After separation, the MACS method yielded a significantly higher percentage of Y-bearing sperm (49.2%) compared to DGC (37.4%), indicating that MACS is a more reliable method for enriching Y-bearing spermatozoa for clinical purposes. In conclusion, MACS appears to be a safe and effective method for sperm selection, consistently producing good results. This technique may improve pregnancy rates when used alongside other sperm selection methods in ICSI therapy.