Cigarette smoking has been reported to induce intrafollicular oxidative stress that may lead to DNA damage. The purpose of this study was to determine damage in DNA in human cumulus cells caused by tobacco smoke in females who had received in vitro fertilization. The level of DNA damage in freshly isolated cumulus cells was determined by comet assay. Statistically significant increase (p<0.05) was observed in damaged nuclear DNA in smokers, both at basal level and after oxidative stress induced by hydrogen-peroxide. Since cumulus cells have an important role in oocyte maturation, ovulation and fertilization, this method could be used both as a test for the evaluation of the biological potential of the female reproductive system and as a direct means to measure certain toxic effects.