The present study addressed the successful in vitro seed germination, bulblet regeneration, increased bulblet size, hardening, and acclimatization of Persian lily (Fritillaria persica). Seed germination rate was recorded as 86.7% and 96.7% after 2 and 3 months of cold treatment at 4 °C, respectively. Bulblet explants taken from a germination experiment were cultured on Murashige and Skoog (MS) medium supplemented with 0.5-2.0 mg/L thidiazuron (TDZ). Maximum bulblet regeneration frequency and bulblets per explants were achieved on a medium containing 2.0 mg/L TDZ. Leaf disc and leaf scale explants isolated from germinated seedlings were also cultured on MS medium containing 2-6 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) with a constant concentration of 0.2 mg/L kinetin. Both explants yielded the highest bulblet regeneration on MS medium containing 2 mg/L 2,4-D and 0.2 mg/L kinetin. A combination of 1.5 g/L agar and 1.6 g/L GELRITE resulted in a maximum bulblet diameter with a higher number of roots. The addition of 10 g/L NaCl to the culture medium resulted in the best root and leaf formation and secondary bulblet regeneration. Exposure of regenerated bulblets to 10 g/L NaCl at 4 °C for 2 months on a medium with 20 g/L sucrose was also found to be beneficial for acclimatization.