Fusarium oxysporum f. sp. cubense (Foc) is the most destructive fungus that causes Fusarium wilt disease, which has led to the loss of some banana germplasm. This research aims to identify banana mutants (Musa acuminata, AAA) resistant to Fusarium wilt using both in vitro selection with culture fluid filtrate (CFF) and in vivo selection methods with conidia of Foc Tropical Race 4 (TR4). Explants from in vitro shoot mutants, derived from gamma irradiation, were cultivated on Murashige and Skoog's media supplemented with increasing concentrations of CFF of Foc, ranging from 40% to 60% (v/v). This study demonstrates that the technique effectively selected banana shoots that were insensitive to CFF from Foc. The presence of CFF as a selective agent reduced the growth of the banana shoots by 13.88% to 48.99%, depending on the gamma irradiation dose from which the variants originated. A correlation analysis revealed a strong negative relationship (R² = 0.70) between CFF concentration and the shoots and nodule-like meristems produced. In vivo selection using a conidial suspension of Foc TR4 at a concentration of 2.5 × 107 mL-1 resulted in 118 plants (23.7%) surviving. Evaluation of banana plants in the greenhouse 24 weeks after infection identified 28 plants (34.6%) from the selected mutans as potentially resistant to Fusarium wilt; most of these plants originated from gamma-irradiated explants at doses of 30 and 50 Gy. This study demonstrates that developing a banana mutant cultivar, 'Ampyang', which is resistant to Fusarium wilt, through gamma irradiation and both in vitro and in vivo selection, is a promising method for identifying resistant plants. However, further assessments via molecular characterization and field evaluations in disease-endemic areas are necessary to confirm the resistance of the obtained mutants.