The aim of this study was to investigate the mechanism of cell activation induced by extremely low frequency magnetic fields (ELF-MF) (50 Hz) in human cells. We examined the production of free radicals in human umbilical cord blood-derived monocytes and in human Mono Mac 6 cells. The release of superoxide radical anions was analyzed using nitroblue tetrazolium chloride and the total of reactive oxygen species (ROS) was detected using dihydrorhodamine 123. Our results show a significant increase of superoxide radical anion production up-to 1.4 fold as well as an increase in ROS release up-to 1.2 fold upon exposure of monocytes to 1 mT ELF-MF (45 min). Mono Mac 6 cells exhibit higher superoxide radical anion and ROS production up-to 1.4 and 1.5 fold, respectively. These results indicate that Mono Mac 6 cells are more sensitive to ELF-MF than monocytes. Using diphenyleneiodonium chloride (DPI) a specific inhibitor for the NADPH oxidase, the MF-effect was not inhibited in Mono Mac 6 cells. Therefore, we suggest that ELF-MF exposure induces the activation of NADH oxidase in these cells. However, the MF-effect was inhibited by DPI in monocytes, indicating the activation of the NADPH oxidase after exposure to ELF-MF.