Cellular imbalance in the levels of antioxidants and reactive oxygen species (ROS) is directly associated with a number of pathological states and results in programmed cell death or apoptosis. We demonstrate the use of in vitro cultured Spodoptera frugiperda (Sf9) insect cells as a model to study oxidative stress induced programmed cell death. Apoptosis of in vitro cultured Sf9 cells was induced by the exogenous treatment of H2O2 to cells growing in culture. The AD50 (concentration of H22 inducing about 50% apoptotic response) varied with the duration of treatment, batch to batch variation of H2O2 and the physiological state of cells. At 24 h post- treatment with H2O2. AD50 was about 475 μm. Apoptosis could also be induced by in situ generation of H2O2 by the inhibition of catalase activity upon hydroxylamine treatment. Hydroxylamine acted synergistically with H2O2 with an AD50 of 2.2 mM. DMSO, a free radical scavenger, inhibited H2O2-induced apoptosis thereby confirming the involvement of reactive oxygen species. Exposure of cells to UV radiation (312 nm) resulted in a dose-dependent induction of apoptosis. These results provide evidence on the novel use of insect cells as a model for oxidative stress-induced apoptosis.