Abstract: This study aimed to establish a protocol for the micropropagation of the cubiu (Solanum sessiliflorum) varieties Santa Luzia (SL) and Thaís (TH). Firstly, nodal segments were cultured onto MS-based medium supplemented with vitamins, mio-inositol (200 mg L-1), glycine (400 mg L-1), sucrose (30 g L-1) and agar (6.5 g L-1), plus NAA (0, 0.25 and 0.5 mg L-1) and BAP (0, 1.0, 2.0 and 3.0 mg L-1). In the second experiment, nodal segments were placed on MS medium, plus vitamins, sucrose (15 g L-1), agar (6.5 g L-1) and IAA (0 and 0.01 mg L-1) in combination with KIN, TDZ or ZEA at 0, 0.64, 1.25, 2.50, 5.0, 10.0 and 20.0 mg L-1. The combinations of NAA and BAP inhibited shoot-bud growth and maintenance of the apical dominance. High TDZ concentrations induced prolific shoot formation with the greatest number of axillary shoots, although with indefinite morphology in many cases. The combination of 0.01 mg L-1 IAA and 5.0 mg L-1 TDZ induced the most intense axillary shoot proliferation in SL, whereas 2.5 mg L-1 TDZ and 0.01 mg L-1 IAA, combined with 1.25 mL-1 TDZ, were most effective for adventitious shoot-bud differentiation. For TH, the proliferation of axillary and adventitious buds was improved in the treatment with 5.0 mg L-1 TDZ and 2.5 mg L-1 TDZ, respectively. After 60 days of cultivation, the adventitious and axillary shoot-buds on MS-based medium lacking growth regulators were 4.2±0.32 cm long and the plants presented a well-developed root system.