Cerium dioxide nanoparticles (CeO 2 NP) are widely used for industrial purposes, as in diesel, paint, wood stain and as potential therapeutic applications. The Organization for Economic Cooperation and Development included CeO 2 NP in the priority list of nanomaterials requiring urgent evaluation. As metal nanoparticles can cross the blood-testis barrier, CeO 2 NP could interact with spermatozoa. The genotoxicity of CeO 2 NP was demonstrated in vitro on human cell lines and mouse gametes. However, the effects of CeO 2 NP on human spermatozoa DNA remain unknown. We showed significant DNA damage induced in vitro by CeO 2 NP on human spermatozoa using Comet assay. The genotoxicity was inversely proportional to the concentration (0.01 to 10 mg·L −1). TEM showed no internalization of CeO 2 NP into the spermatozoa. This study shows for the first time that in vitro exposure to very low concentrations of cerium dioxide nanoparticles can induce significant DNA damage in human spermatozoa. These results add new and important insights regarding the reproductive toxicity of priority nanomaterials, which require urgent evaluation.