The aim of this study was to observe responses of two banana cultivars ‘Ambon' and ‘Raja Bulu' on different BA concentrations and effects of different media and fertilizer on survival and growth of plantlets. Sterilized explants were cultured on initiation medium (MS with 1mg L-1 BA) for 4 weeks, then subjected to media MS with 2.5, 5.0, or 7.5 mg L-1 BA. Numbers of shoot buds, shoots and propagules were recorded after 4 consecutive passages with 4 weeks intervals. Rooted plantlets were acclimatized in three different media, then treated with or without NPK (32:10:10) fertilizer solution once a week. After 2 months, the survival and growth of plantlets were recorded. Cultures of banana ‘Ambon Kuning' showed higher regenerative capacity compared to ‘Raja Bulu', producing higher numbers of shoot buds, shoots and propagules. The best medium for propagule proliferation of both banana cultivars was MS+5 mg L-1 BA, producing 40.7 propagules for ‘Ambon Kuning', and 12.3 propagules for ‘Raja Bulu' per explant. In all acclimatization media tested, 100% of plantlet survival was achieved. The best plantlet growth was found in sand: compost (1:1,v/v) with application of NPK solutions. The in vitro-derived plants were planted in the field and produced fruits of high quality.