Objective To compare different culture media for their effects on sperm motility and lipid peroxidation. Design The ability of four different culture media to sustain the motility of human spermatozoa during an overnight incubation was examined in relation to the induction of lipid peroxidation. The role of transition metals in the genesis of peroxidative damage was investigated in experiments involving the addition of iron or the chelating reagent ethylenediamine tetra acetic acid (EDTA). Setting Academic research institute. Main Outcome Measures The generation of malondialdehyde and 4-hydroxyalkenals after the addition of a ferrous ion promoter and the measurement of sperm motility. Results Spermatozoa incubated in Ham's F-10 medium (GIBCO, Paisley, Scotland) exhibited a marked loss of motility in association with a significant increase in peroxidative damage. Addition of EDTA to Ham's F-10 medium significantly reduced the degree of lipid peroxidation and induced a simultaneous increase in sperm motility. Conclusions Ham's F-10 medium is a suboptimal medium for the long-term culture of human spermatozoa because it induces peroxidative damage and a concomitant decline of sperm motility.