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Abstract
Three ml of semen collected from 20 volunteers were divided into three aliquots: a) semen cryopreserved for 48 hr then swim-up method; b) using swim-up method then cryopreserving for 48 h; and c) using 45%–90% two layers PureSperm to prepare sperm then cryopreserved for 48 h. Motility characteristics were evaluated before cryopreservation and after thawing. NO was measured using the chemiluminescence method after centrifugation. The recovery rate and motion parameters were the highest in the swim-up group. NO produced in the swim-up group was significantly lower than the PureSperm group, and the PureSperm group was lower than the semen treated with cryoprotectant group. Best semen quality was achieved due to the decreased NO production.