Traditional asexual propagation of hops limits its phytosanitary quality and the concomitant plant availability. This scenario worsens with the global climate change. In this study we develop protocols for in vitro propagation of Humulus lupulus var. mapuche. Two explant types were evaluated: shoot tips, from field material collected in Río Negro province (Argentina), and nodal segments, from vitroplants. Different growth regulator, such as 6-Benzylaminopurine, gibberellic acid, indole-3-butyric acid and indole-3-acetic acid, was used to induce desired morphogenic responses. Additionally, the impact of abscisic acid application on acclimatization was assessed, and resulted in robust growth and reduced plant stress symptoms during ex vitro adaptation. Also, the study develops a protocol for synthetic seeds production, with storage at 4°C to enhance conservation and plant material transport, ensuring a constant supply of female plants. This comprehensive approach addresses current producer demands and emerging climate challenges. The findings highlight the fundamental role of in vitro multiplication of hops in improving plant material availability and quality, with significant implications for the brewing industry and related sectors.