This study investigated the antioxidant potential of callus cultures derived from the endangered medicinal plant Ferula tadshikorum Pimenov (Apiaceae Lindl.), an endemic species in Central Asia. The plant’s long life cycle and overharvesting threaten its survival, necessitating the development of in vitro microclonal propagation protocols as a sustainable solution for bioactive compound production. Two propagation methods, indirect organogenesis and indirect somatic embryogenesis, were optimized using zygotic embryos as explants. Callus cultures at early stages, cultivated on Murashige and Skoog medium supplemented with 0.5 mg/l 2,4-dichlorophenoxyacetic acid, demonstrated antioxidant activity, with 2,2-diphenyl-1-picrylhydrazyl (DPPH) half-maximal inhibitory concentration (IC₅₀) values as low as 15.41 µg/mL, comparable to ascorbic acid (15.17 µg/mL), while the radical-scavenging activity in the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay was even higher. Optimal bioactivity and extract yields were achieved on a Murashige and Skoog medium supplemented with 2.4-dichlorophenoxyacetic acid (0.5 mg/l) and Kinetin or Thidiazuron. Statistical analysis confirmed the influence of hormonal treatments on antioxidant properties. These findings highlight the dual potential of in vitro propagation for conserving F. tadshikorum and providing a sustainable source of bioactive compounds with significant antioxidant properties, supporting pharmaceutical applications. However, challenges in translating these findings into large-scale production or commercial applications must be addressed, including the optimization of protocols for industrial scalability and cost-effectiveness.