<p>In recent years, research on <i>Cannabis sativa</i> L. has undergone exponential growth, driven by the adoption of more permissive regulatory frameworks for the species in many regions, notably in North America. In this rapidly evolving scientific and applied context, significant advances have been made in plant biotechnology, focusing on the development and optimization of in vitro culture protocols for genetic improvement and clonal propagation of cannabis. The introduction of next‑generation plant growth regulators and the refinement of innovative culture‑media formulations, together with the implementation of advanced acclimatization strategies, have enabled researchers to overcome challenges such as genotype‑dependent recalcitrance, low tissue‑differentiation efficiency, and nutritional imbalances commonly encountered in traditional procedures. Micropropagation, known as a technique for large‑scale multiplication and long-term conservation of elite germplasm, offers critical advantages including the production of pathogen‑free plantlets, guarantees genetic uniformity, and the efficient use of cultivation space for the exploration of agronomic traits and phytotherapeutic phenotypes. This review provides a critical and detailed analysis of the current state of the art, emphasizing recent innovations in culture‑media formulations and micropropagation protocols specifically tailored for <i>C. sativa</i>, with the aim of outlining future perspectives and potential industrial applications.</p>