Leaf discs, internodal cuttings and root cuttings were taken from healthy lavender plants( Lavandula officinalis L.), and were cultured on different medium types(Murashige and Skoog,Nitsch&Nitsch and modified Murashige and Skoog). Different cold pre-treatments (3°C for 1, 3, 5 &7 days ) ,different auxin types (NAA, 2,4-D and I.B.A.), and different 2,4-D concentrations(0.0, 0.5, 1.0, 2.0 &4.0mg/L ) were evaluated.Also,different cytokinin types(Kin, BAP and 2-ip), Different BAP. Concentrations (0.0, 0.5, 1.0, 2.0 and 4.0mg/L) and combinations between both auxin &cytokinin were considered to maximize callus formation &oil yielding ability. The produced global or callus were subjected to maturation and treated with either Glycine,Yeast extract or Tryptophan treatments.In the same time different concentrations(0.0,50,100,150,and 200 mg/l to maximize oil content and oil characters. It was found that modified MS medium ,7 days cold pretreatment,2.0 mg/l 2,4-D. &0.5 mg/l BAP.and 100 mg/L induced the best results. The volatile oil composition of the four treatments of organic additives i.e. Glycine, Yeast extract,and Tryptophane were Twenty compounds. Comparison of the analytical data of the oils revealed marked differences in qualitative and quantitative composition.especially Linalool, camphor,and β-caryophyllene. Keywords:-Lavender, growth regulators, Additives, Oil content, in vitro.