Dynamic assessment of human sperm DNA damage II: the effect of sperm concentration adjustment during processing

To evaluate the effect of sperm concentration adjustment in human ejaculates on the sperm DNA quality and longevity.

Semen samples were obtained from 30 donors with a normal spermiogram. Following centrifugation, the sperm pellet was resuspended in PBS, and the sperm concentration adjusted to 200, 100, 50, 25, 12, and 6 × 10⁶/mL. Each set of samples was incubated at 37 °C for 24 h, and the sperm DNA damage was assessed using the chromatin-dispersion test following 0 h, 2 h, 6 h, and 24 h of incubation.

Sperm DNA fragmentation (SDF) did not differ between the selected experimental conditions at T0; however, Kaplan-Meier estimates for survival showed significant differences with respect to the dilution and time (all P values were smaller than .001). DNA fragmentation in semen samples adjusted to 200 × 10⁶/mL was approximately 3.3 times higher when compared to samples containing 25 × 10⁶/mL and 3.9 higher in comparison with samples adjusted to 12 × 10⁶/mL following 2 h of in vitro incubation. Although there was evidence of individual variation in SDF during the incubation period, the general finding was that lower sperm concentrations resulted in a slower rate of DNA fragmentation.

Incubation of spermatozoa for ART purposes should be done following a concentration adjustment below 25 × 10⁶/mL in order to avoid a higher susceptibility of the sperm DNA molecule towards fragmentation.