This investigation was carried out at the Tissue Culture Lab., Hort. Dept., Fac. Agric., Zagazig Univ., Egypt during the two successive seasons of 2014 and 2015 to produce transplants of two tomato hybrids through tissue culture technique. A protocol was developed for shoot multiplication and rooting in two tomato (Lycopersicon esculentum Mill.) hybrids i.e., Silla 108 and Farrah. Shoot tips were cultured into Murashige and Skoog media (MS medium) with different concentrations of benzyl adenine (BA) and Kinetin (Kin) 1.5 mg BA/l, 1.5 mg BA/l + 1 mg Kinetin/l, 3 mg Kin/l and 4 mg Kin/l) compared with MS basal medium as a control treatment, the obtained results illustrated that supplementing BA at 1.5 mg/l gave the highest value of number of shoots/explant of Sila 108 hybrid followed by kinetin at 4 mg/l for Farrah hybrid in both seasons. Multiplicated shoots of two tomato hybrids were excised and cultured on MS medium supplemented with different rooting growth regulators including indole acetic acid (IAA) at 0.5 and 1 mg/l, Naphthalene acetic acid (NAA) at 1 mg/l and indole butyric acid (IBA) at 1mg/l compared to the control treatment (without growth regulators). From the foregoing results, it could be concluded that supplementing IAA at 1 mg/l and NAA at 1 mg/l to MS media were the best treatment for rooting stage of Farrah and Sila 108 tomato hybrids. The plantlets were transferred to plastic cups containing peatmoss + sand at 1:1, peatmoos + vermiculite at 1:1 (V/V) or peatmoss alone. It could be concluded that the agriculture media for acclimatization of plantlets of Farrah and Sila 108 tomato hybrids produced from tissue culture technique were peatmoss and peatmoss+ vermiculite at 1:1 (V/V).