An investigation on Lycopersicon esculentus cvs. Castle Rock (CR), Strain B and Super-Marmand (Tomato) was conducted to standardize the method of high frequency regeneration. Cotyledon and hypocotyl from 12 days old seedlings were used. Comparison was made on cotyledon and hypocotyl explants cultivated on 4 different regeneration media of three cultivars. The best results for shoot primordial numbers per explants were observed of hypo-cotyledon explants with MS medium containing 1mg/L BA of cultivars Castle Rock and Super-Marmand, was reached (91-87% respectively), while the best results for shoot primordial numbers per explants reached the highest value (83%) for strain B by using MS medium supplemented with 0.5 mg/L BA compared to the other treatments. While, the best results for shoot primordial numbers per explants were observed of cotyledon explants with MS media containing 2.0 mg/L BA of cultivars Castle Rock, strain B and Super-Marmand compared to the other treatments, was reached (79,72 and 80%, respectively). The percentage of shoots forming roots was reached the highest value 95 and 90 % of Castle Rock (CR) and Super-Marmand, respectively by using MS medium supplemented with 1.0 mg/ L IBA, while the best results of shoots forming roots was reached the highest value (92%) for strain B by using MS medium supplemented with 0.5 mg/L IBA and 0.5 mg/L NAA. Tomato plants are able to regenerate within 2 months and the young plantlets can be transferred to pots for hardening. Hardening procedure for tissue culture raised plantlets was standardized using sterile mixture of sand and soil in plastic pots.