The olive (Olea europea L.) explants of Itrana cultivar were cultured on different types of medium i.e; Olive medium (OM), Woody Plant medium (WPM) and Murashige & Skoog medium (MS). Also, different explant types (shoot tips and nodal cuttings) were investigated. The results showed that the Olive medium (OM) with nodal cuttings proved to be the best medium and explant for establishment of olive plants cv. Itrana. The best multiplication for cv. Itrana was obtained with culturing on OM medium supplemented 3 mg l^-1 zeatin. Shoot elongation was carried out on OM medium containing different concentrations of Gibberellic acid (GA₃). Adding 2 mg l^-1 of GA₃ to the culture medium proved to be the best concentration for obtaining the highest shoots (10.66 cm). Shoot rooting was carried out on half strength OM medium with different types of auxins (IBA, NAA or IAA). IBA proved to be the most effective one for inducing rooting of shoots. Adding 3 mg l^-1 of IBA achieved the highest number of roots (2.66) and root length (3.66 cm) and it was proved to be the best concentration for rooting stage. Adding 0.5 mgl^-1 of Putrescine to the rooting medium proved to be the best concentration for root formation and growth. Finally hardening the produced plantlets were performed successfully in greenhouse in pots containing mixture of peat-moss, vermiculite and sand (1:1:1) V/V/V.