The objective of the research was to determine a protocol for disinfection and in vitro establishment of explants from strawberry crown buds Fragaria x ananassa Var. Aroma, from the Minor Town Center Mallampampa, Huancabamba and the La Florida sector, Chontabamba district, Oxapampa, Pasco, Peru, through a randomized complete block design. Three disinfection methods were used T1 (adapted from Coba, 2009), T2 (adapted from Sánchez-Cuevas and Salaverría, 2004)and T3 (adapted from Mamani, 2005) and in the establishment were the treatments, t1 (MS+ BAP 0.5 mg l -1+ NAA 0.5 mg l-1), t2 (MS+BAP 0.5 mg l-1+ NAA 0.3 mg l-1+ GA3 0.2 mg l-1), t3 (MS+BAP 0.5 mg l-1+ IAA 0.6 mg l-1), t4 (MS+BAP 0.5 mg l-1+ IAA 0.5 mg l-1+ GA3 0.3 mg l-1) and t5 (MS without regulators). At 36 days after inoculation (dai),the best disinfection treatment was T3, because it presented less contamination by bacteria and less phenolization and without significant differences in fungi and explant survival. The best establishment medium was t3, for present a lower degree of phenolization, greater survival, sprouting and number of shoots and leaves.