Environmental Exposure of Sperm Sex-Chromosomes: A Gender Selection Technique

Preconceptual sex selection is still a highly debatable process whereby X- and Y-chromosome-bearing spermatozoa are isolated prior to fertilization of the oocyte. Although various separation techniques are available, none can guarantee 100% accuracy. The aim of this study was to separate X- and Y-chromosome- bearing spermatozoa using methods based on the viability difference between the X- and Y-chromosome-bearing spermatozoa. A total of 18 experimental semen samples were used, written consent was obtained from all donors and results were analysed in a blinded fashion. Spermatozoa were exposed to different pH values (5.5, 6.5, 7.5, 8.5, and 9.5), increased temperatures (37 degrees C, 41 degrees C, and 45 degrees C) and ROS level (50 mu M, 750 mu M, and 1,000 mu M). The live and dead cell separation was done through a modified swim-up technique. Changes in the sex-chromosome ratio of samples were established by double-label fluorescent in situ hybridization (FISH) before and after processing. The results indicated successful enrichment of Xchromosome- bearing spermatozoa upon incubation in acidic media, increased temperatures, and elevated H2O2. This study demonstrated the potential role for exploring the physiological differences between X- and Y-chromosome-bearing spermatozoa in the development of preconceptual gender selection.