The influence of pulsed electric field (PEF) (0.5-2 kV/cm) treatment of wheatgrass (Triticum aestivum L.) seeds, with different water contents, on antioxidant metabolism in the resultant seedlings was investigated. Imbibing seeds to a water content of 45% or greater prior to PEF treatment increased the glutathione level and activities of superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase and ascorbate peroxidase in the resultant seedlings, compared to untreated controls. Pre-culture of human intestinal Caco-2 cells with simulated gastrointestinal digests of electrostimulated seedlings enhanced the ability of Caco-2 cells to cope with H2O2-induced oxidative damage, determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) release assays. The Caco-2 cell MTT and LDH assays correlated better with the increases in seedling glutathione content and antioxidant enzyme activities compared to the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) total antioxidant capacity assay, an assay commonly used to determine the ability of plant extracts to protect cells from oxidative damage. These results demonstrate for the first time that PEF treatment of imbibed seeds can stimulate changes in metabolism in the resultant seedlings, increasing the bioprotective potential of their shoots/sprouts and hence value as functional foods.