This study investigated the effect of silymarin on human sperm quality during cryopreservation. Samples were collected from 20 normospermic individuals, and each sample was divided into different concentrations of silymarin comprising the following groups: (0, 20, 100, 500, and 1000 mg/mL silymarin). Sperm quality parameters, such as plasma membrane integrity, mitochondrial membrane potential, acrosomal membrane integrity, and caspase 3 were estimated. Silymarin concentrations of 100–500 mg/mL significantly increased motility, plasma membrane integrity, and mitochondrial activity compared with the frozen control group. Acrosomal integrity was increased in the 1000 mg/mL silymarin group. Moreover, 20 and 100 mg/mL concentrations significantly decreased the percentage of caspase 3. The addition of silymarin antioxidant to the frozen medium reduced damage in the sperm after freezing and thawing. This is the first study that showed silymarin can be useful in cryopreservation of human sperm.