Fig 1.
CONSORT flow diagram.
Fig 2.
For the study cohort, ejaculates were processed by a multilayer density gradient (Enhance-S Plus Cell Isolation Media, 100%; Vitrolife, Goteborg, Sweden) comprised of four concentrations: 20%, 40%, 60%, and 90%. After removing the seminal fluid, the resuspended sperm pellet was layered on top, and spermatozoa were able to self-select for 90 minutes at 4°C. Y-bearing spermatozoa were retrieved from the upper, 20% layer, and heavier X-bearing spermatozoa were retrieved from the bottom, 90% layer.
Fig 3.
A total of 1,317 couples undergoing ICSI cycles with preimplantation genetic testing were included in our study. Couples (n = 1,212) who did not desire a specific offspring sex underwent ICSI cycles with PGT-A solely for aneuploidy, while the remaining 105 elected to undergo ICSI with PGT-A for sex selection.
Table 1.
Demographics and semen parameters for the control and study groups according to sex.
Table 2.
Proportion of sperm cells enrichment and resulting embryos for female and male sex.
Fig 4.
For couples selecting for female sex, our processing method was able to yield an increased proportion of X-bearing spermatozoa (left). Similarly, our method yielded a greater proportion of Y-bearing spermatozoa for couples selecting for male sex (right).
Table 3.
ICSI outcomes by sex preference.
Table 4.
Ploidy of tested embryos for the control and study groups according to sex.