Populations of Agave potatorum Zucc. have been overexploited from their habitat for the commercial production of mezcal, a traditional Mexican liquor. For this species, micropropagation is the only method for cloning selected genotypes. The aim of this study was to establish an efficient protocol for the in-vitro propagation of A. potatorum by use of individual shoots. In the propagation stage, the interaction between different concentrations of the cytokinin benzylaminopurine (BAP: 0, 1.5, and 3.0 mg L–1)and the auxin indole acetic acid (IAA: 0, 1.5, and 3.0 mg L–1) was evaluated. For in-vitro rooting, the interaction between different auxin concentrations, namely naphthaleneacetic acid (NAA: 0, 1.5 and 3.0 mg L–1) and IAA (0, 1.5 and 3.0 mg L–1) was evaluated. In the propagation stage, the highest number of shoots was obtained with the combinations of 3.0 mg L–1 BAP + 3.0 mg L–1 IAA, 1.5 mg L–1 BAP + 3.0 mg L–1 IAA, and 3.0 mg L–1 BAP + 1.5 mg L–1 IAA, which yielded 9.87, 9.73, and 9.73 shoots per explant, respectively.  In the rooting stage, the best shoot development was observed in the control treatment and when only 3.0 mg L–1 IAA was supplemented. Finally, after the rooting stage, the plantlets obtained were acclimatized and grown in the field, yielding a survival rate of 98-100%. In conclusion, this propagation protocol contributes to obtaining commercial propagules suitable for establishment in the field.