The present study aimed to investigate the effect of TDZ and BAP on in vitro microtuberization in potato cv. Spunta. Different concentrations (4, 6, 8, and 10 mg/l) of BAP either alone or in combination with three concentrations (1, 2, and 3 mg/l) of TDZ were used for investigating their influence on microtuberization. The results showed that the individual treatment of BAP at the high level (10 mg/l) recorded the highest value of the average number of the produced microtubers/jar (7.1 microtubers/jar) compared to the other treatments (4, 6, and 8 mg/l). The combinational treatments of BAP and TDZ exhibited an increase compared to the individual corresponding concentration of BAP and this increase reached a significant level with the concentrations (4, 6, and 8 mg/l) of BAP, while a slight increase was recorded with 10 mg/l. Furthermore, the combinational treatment of BAP (6 mg/l) and TDZ (1 mg/l) resulted in the highest average number of the produced microtubers/jar (8.1 microtubers/jar) which represents about 41.1 % increase over the individual treatment of BAP (6 mg/l) with the highest average weight of microtuber (116.94 mg) and the highest average diameter of microtuber (5.3 mm). These results confirm the positive effect of the combinational treatment of BAP and TDZ on enhancing microtuber formation in potato cv. Spunta. Therefore, we report here that an efficient protocol for microtuberization in potato cv. Spunta was established. In addition, these promising results are important for developing advanced technology for microtubers mass production in Egypt.