Introduction: Sesamum indicum L. belongs to Pedaliaceae. This family has 60 species. Ethiopia is probably the origin of SESAME. SESAME is one of the important oil seed, industrial and medicinal plant.Aim: Gain sterile seedling of SESAME seed, successfully culture of different part of seedlings, determined the best medium for call genesis and organogenesis and compared the growth’ regulating agents for callogenesis and organogenesis.Material and Methods: In this research, in vitro culture of Sesamum indicum L. was investigated in MS media with different hormones such as IAA, Kin, and NAA. Different part of seedlings grown on hormone free such as root leaf, stem, apical meristem were used as explants. Accidentally locks with 3 repeats and I each epeats 4 samples was used. Results: The best media for callogenesis , with considering the size and stability were mediums supplementedwith10 mg/l IAA/2mg/l Kin, 10mg/l IAA/5mg/lKin, 10mg/l NAA/2mg /lKin, 10mg/l NAA/5mg/lKin, 20mg/l NAA/10mg/lKin. Between the explants that used, apical meristem in medium supplemented with 2mg/l IAA/10mg/l Kin after callogenesis and organogenesis, had a highest means of regenerated plantlets, this plantlets have must resistantly in culture medium and in nature in comparison with plantlets from apical meristem in the medium with different concentration of IAA/Kin and NAA/Kin. Leaf explants, also in medium supplemented with 10mg/l IAA/20mg/l Kin, formed complete plantlets. There is no organogenesis in calli from stem and root explants.Conclusion: In different part of seedlings, apical meristem was the best part for plantlets regeneration and MS medium supplemented with 2IAA/10Kin (mgl-1) was the best medium for plantlets regeneration from apical meristem.