For in vitro propagation of Lilium longiflorum (Meristem and twin scales showed different responses toward different type of media. The best shoot formation response was obtained on Murashige and Skoog medium supplemented with 2.0 mg/l of BAP. At this concentration 84% shoot formation response was obtained after 12.2 days of inoculation. When MS medium was supplemented with different concentrations of auxin and cytokinins the best shoot initiation response was gained at 2.5 mg/l of BAP with 0.5 mg/l of NAA i.e. 88% shoot initiation was obtained after 21 days of explants inoculation. Maximum shoot multiplication response was obtained at 2.5 mg/l of BAP i.e. 4.6 shoots per culture were observed after 15 days of inoculation. MS medium supplemented with 1.5 mg/l of NAA was proved to be optimum for rooting i.e. an average of 5.2 roots were formed after 12.2 days of inoculation. Temperature of 26°C and photoperiod 16 hours & 8 hours dark in 24 hour cycle induced the best response for in vitro shoot formation. Well developed in vitro plants were shifted to pots containing different media including soil, peat, soil + sand and soil+ sand + peat. 100% hardening response was obtained in peat.