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Growing media is one of the important factors that determine the success of the acclimatization of plantlets from in vitro culture. Several media combinations were tried in this study with the aim of 1) Knowing the effect of media combinations on the success of plantlet acclimatization and seedling growth at the enlargement stage, 2) Getting the best media combination for the successful acclimation and enlargement of banana seedlings, and 3) Seeing the possible use of media husk in acclimatization and enlargement of banana seedlings as a result of tissue culture. The study was conducted at the Lampung State Polytechnic net home from May to August 2019. The research was carried out in a Randomized Block Design. Six media combinations were tried, namely: P1: Topsoil + Husk (1: 1); P2: Top Soil + husk (1: 2); P3: Topsoil + husk charcoal (1: 1); P4: Topsoil + Husk charcoal (1: 2); P5: Top Soil + compost (1: 1); P6: Topsoil + compost (1: 2). In acclimatization (planting until the plantlet is 4 weeks old) each treatment planted 30 plantlets, each with 5 replications, while on the enlargement of the seeds each treatment was tried with 15 replications. Observed variables included: Seed height, leaf width, number of leaves, and wet weight. Data were analyzed by analysis of variance and continued with the BNJ test at a 5% level. The results showed: 1) the combination treatment of media had no effect on the percentage of plantlets that grew, and the growth of the number of seedling leaves at the enlargement stage, but markedly affected the growth of seedling height, leaf width, and seedling wet weight, 2) the best combination of media for success plantlet acclimatization and seedling growth rate (seedling height and wet weight) at the enlargement stage are P5 media (Topsoil + 1: 1 compost), then followed by P6 media (Topsoil + Compost 1: 2), and 3). Use of husk media for additional acclimatization media, it can still be used with the success rate of plantlets still reaching 70%, but it is not good to be used at the stage of seedling enlargement until it is ready for planting into the field.