The effects of plant growth regulators and explants on callus induction were investigated for the purpose of developing a protocol for callus induction of garden cress (L. sativum. L). Three explants namely, leaves, hypocotyls and roots obtained from 7 days -old in vitro germinated seedlings were cultured on MS medium supplemented with different concentrations of 2,4-dichloro-phenoxyacetic acid (2,4-D) and α-naphthalene acetic acid (NAA). The highest callus weight (2.13) gm was obtained when leaves explant was cultured on MS medium containing 2.0 mg/L 2,4-D. Addition of cytokinns Benzyl adenine (BA) and Thiadizuroun (TDZ) enhanced the callus weight as well as callus growth index. The maximum callus weight obtained when leaves explant cultured on MS media suplemented with 2.0 mg/L 2, 4-D in combination with 3.0 mg/L BA. This procedure can be advantageously of extraction of active ingredient as well as developing of in vitro regeneration protocol that can use in genetic improvement of this multipurpose medicinal plant.