Dipteryx alata Vog. (barueiro) is a native fruit of the Cerrado used in food and medicine. The species has never been domesticated and is currently at risk of extinction. Plant tissue culture is an important tool for the conservation of germplasm and for quick, large-scale propagation of quality seedlings. While, this technique has never been used with barueiro, it could be a valuable tool in the domestication of this species. The objective of this work is to evaluate concentrations of MS medium salts (25, 50, 75 and 100%) supplemented with coconut water (100 mL L -1 ), dwarf banana pulp (60 g L -1 ) or the absence of any supplement in the in vitro establishment of barueiro seeds (Dipteryx alata Vog). The experiment used a completely randomized design (CRD) with a 3x4 factorial scheme (three cultivation media types and four concentrations of MS salts) and three repetitions. The following characteristics were evaluated: germination rate, shoot length , largest root length , stem diameter , number of leaves , chlorophyll A , chlorophyll B , total chlorophyll, and the fresh and dry mass of the seedlings. The coconut water and banana pulp supplements did not improve the in vitro development of the barueiro seedlings and the full concentration of MS salts did not improve the initial development of the seedlings. However, the pure MS medium at a concentration of 25% MS salts was the best option for the in vitro establishment of Dipteyx alata.