Background: Human semen cryopreservation considered as indispensable part of assisted reproductive centers, androl- ogy laboratories and sperm bank. This technique is associated with significant damage to sperm. Adding an antioxidant to the cryopreservation medium could effectively mitigate these ad- verse effects. Lycopene, a naturally occurring red carotenoid with potent antioxidant properties, has been shown beneficial effects on various sperm parameters. This investigation studied the effect of lycopene supplementation on Acrosome Reaction, mitochondrial membrane potential (MMP) and DNA fragmen- tation during the cryopreservation of semen samples from as- thenozoospermic men. Materials and Methods: Semen samples were collected from 30 asthenozoospermic men. Each sample was then divided into three groups: Control (fresh), Freeze (treated with cryoprotect- ant alone), and Freeze + Lycopene (treated with cryoprotectant and 5 µmol/L lycopene). The samples in the freezing groups were cryopreserved using a human sperm freezing medium and a rapid freezing method. For each sample, acrosome re- action with fluorescein isothiocyanate-labelled pisum sativum agglutinin (FITC-PSA) staining, MMP was evaluated using Rhodamine 123 staining, and DNA fragmentation was exam- ined using SDFA kit. Data were statistically analyzed using the repeated measures analysis method. Results: Our results showed a significant increase in the mean level of sperm DNA fragmentation and also a significant de- crease in the mean percentage of sperm with intact acrosome and sperm mitochondrial membrane potential in the freeze group compared to the control group (P<0.001). While in the freeze + lycopene group, a significant decrease in the mean level of sperm DNA fragmentation (P<0.001) and a significant increase in the mean percentage of sperm with intact acrosome (P=0.003) and mean sperm mitochondrial membrane poten- tial (P<0.001) were observed compared to the freeze group (P<0.001). Conclusion: Based on our results we concluded that lycopene reduces the adverse effects of cryopreservation on sperm qual- ity in asthenozoospermic men.
