Callus of five onion genotypes representing two species. Allium cepa and A. fistulosum, and their interspecific hybrid were used for establishing suspension cultures. Cultures were derived from callus that had been maintained on solid media and routinely subcultured for four years and from callus induced within six months of this experiment. Long-term callus from which plants were routinely regenerated and newly-induced callus were composed of cells which were, for the most-part, meristem-like with higher mitotic indices than cells from long-term callus which had been maintained as callus but had lost us capability to regenerate plants, these cells were large with small nuclei. Callus from newly-induced and long-term regenerable cultures were selected for further studies. Eight liquid media with factorial combinations of plant growth regulators were tested. Cells cultured in BDS liquid medium supplemented with 0.5 mg/l ABA and 1.0 or 2.0 mg/l 2,4-D without e-BA had higher mitotic indices and plant regeneration percentages than did cells cultured in the same media without ABA and with 6-BA. Suspension cultures from A. fistulosum and interspecific hybrids with A. fistulosum produced the highest numbers of plants regenerated.