Analysis of cryopreservation effects on sperm DNA integrity.
Laboratory of Histology–Embryology of medicine faculty, Sfax, Tunisia.
Fifteen semen samples from men undergoing infertility investigation.
Neat semen samples were cryopreserved in liquid nitrogen using a commercial freezing medium (SpermFreeze, Fertipro, Belgium) according to the manufacturer's instructions. Samples were thawed at room temperature.
Sperm DNA fragmentation was assessed using terminal deoxynucleotidyl transferase (Tdt) mediated dUTP nick end labeling and sperm DNA oxidation was determined using a fluorescent assay (OxyDNA test) for the detection of 8-oxoguanine. Evaluation of DNA fragmentation and oxidation rates was carried out before and after cryopreservation using flow cytometry.
A significant decrease in sperm motility and viability was observed after cryostorage. In addition, sperm DNA fragmentation and DNA oxidative damage increased significantly after cryopreservation/thaw.
Cryopreservation has deleterious effects on sperm DNA by inducing DNA fragmentation and oxidation but the mechanisms underlying such damages need to be elucidated by further investigations.