Abstract Passiflora alata Curtis (sweet passion fruit), native to Brazil, is one of the commercially cultivated species of the genus, being consumed in natura due to its sweet taste, and also used as an ornamental. The goal of this work was to develop new strategies for in vitro culture of P. alata , including plant production, and establishment of callogenesis and cell suspension cultures. Shoot elongation rate and number of nodes per shoot of the primary cultures were improved by adding coconut water to the medium. Plants derived from primary cultures were used as sources of nodal, internodal and leaf explants. Shoot formation occurred through distinct pathways in response to 6-benzyladenine. Internodal and leaf explants displayed both direct and indirect organogenesis, resulting in the formation of 9.9 ± 1.3 and 2.0 shoots per explant, respectively. Nodal segments originated organogenic calluses, with the production of 12.9 ± 1.5 shoots per explant. Shoots were transferred to elongation medium (MSM + 10% coconut water), and showed root formation at a frequency of 100%, after 30 days of culture. Friable calluses were induced from nodal and leaf segments in the presence of PIC, and those obtained from leaves on medium supplemented with 28.9 μM PIC were selected for the establishment of cell suspension cultures.