Saffron (Crocus sativus L.) is the most expensive spice in the world. This plant species is propagated vegetatively by the formation of daughter corms from the mother plant. Unfortunately, many factors prevent the efficient propagation of saffron through this traditional practice. For example, the low production rate of daughter corms and the risk of fungal contamination affect the quality of daughter corms. Therefore, the traditional propagation method cannot meet the demand for planting material. The use of biotechnological tools, especially in vitro culture techniques, can be of great benefit in propagating saffron. Therefore, in recent years, various regeneration systems have been established for saffron through somatic embryogenesis and organogenesis using different explant types, media components, plant growth regulators (PGRs) and culture conditions. In vitro culture methods allow obtaining large amounts of propagation material for the saffron plant in a short time. This research was conducted to reveal the effects of different nutrient medium and hormone combinations on the in vitro propagation of saffron and to establish a sustainable in vitro micropropagation protocol. As a result of the research, it was revealed that the use of relatively high amounts of BAP in addition to maintaining a high cytokinin/auxin ratio in basic media (MS or DKW) is necessary for a sustainable saffron in vitro micropropagation protocol.