Abstract: The aim of this study was to develop a shoot regeneration protocol for Tectona grandis by examining the effects of pretreatments, regeneration culture medium, adventitious rooting, and acclimatization. The best organogenic potential was achieved when hypocotyls and cotyledonary segments from seedlings germinated on a medium containing TDZ, were cultured for four weeks on MS medium supplemented with BAP/GA3. With TDZ pretreatment during seed germination, up to 70% of the cotyledonary segments (0.5 TDZ pretreatment and 1.0 BAP + 0.5 GA3 during regeneration, mg L-1) and 60% of the hypocotyl explants (0.1 TDZ pretreatment and 1.0 BAP + 0.5 GA3 during regeneration, mg L-1) resulted in shoot regeneration. Finally, 65% of the regenerated shoots rooted, of which 60% were successfully acclimatized ex vitro. Our protocol increases regeneration efficiency for teak and it can serve as a platform for genetic transformation to improve this important tree species.