The study has aimed to investigate the effect of Quercetin, as a potent antioxidant, on preventing the negative effects of freezing process on sperm quality of patients with Asthenospermia. Semen sample from 25 patients was randomly divided into three groups; fresh, control and Quercetin (50 μM). Seven days after freezing, sam- ples were thawed at ambient temperature. Total motility, progressive sperm motility, normal morphology, viability and DNA integrity were evaluated according to WHO criteria, Papanicolaou, eosin- nigrosine and acridine orange staining respectively. In addition, the health of sperm membrane and mitochondrial membrane potential was assessed by HOS test and rhodamine staining. MDA and antioxidant enzyme activity were also evaluated using ELISA method. In contrast to the fresh group, the mean level of MDA and DNA damage had significant increase in the control group and decreased significantly sperm quality (p ≤ 0.001). The mean percentage of total mo- tility and progressive motility, normal morphology, viability and antioxidant enzyme activity had significant increase in the Quercetin group than the control group. In the Quercetin group, the MDA level and DNA damage also had significant reduction in comparison with the control group (p ≤ 0.001). Therefore, the Quercetin supplemen- tation improves the quality of cryopreserved human semen.