Human diploid fibroblasts (strain VH-10) were exposed to the direct-acting alkylating agent, ethylene oxide (EtO), in vitro, and the frequency of HPRT mutants was evaluated by selection in medium containing 6-thioguanine. A dose-dependent increase of the mutant frequency was found in the dose range of 2.5-10 mMh of EtO. The EtO-induced mutant frequency increased 5-19 times the background frequency at low or moderately toxic doses, which indicates that EtO is a strong mutagen in human fibroblasts in vitro. The mutagenic potency was 9.8 x 10(-6) per mMh.