An efficient and reproducible method for regeneration of squash (Cucurbita pepo L.) plants via somatic embryogenesis was developed. The influence of genotype, explant source and growth regulators concentrations on somatic embryogenesis induction was investigated. Embryogenic callus was induced from different organs of two squash genotypes. The embryogenic callus was developed within 13- 20 weeks incubation on MS medium containing different plant growth regulators combinations of auxin and cytokinin. Induction of embryogenesis in different explants ranged from 7 to 100 % depending on the organ and genotype used. Successful acclimatization of squash in vitro plants was achieved in the greenhouse and field. Regenerated plants appeared morphologically normal and set flowers and fruits with seeds that could germinated normally.