Callus cultures of Withania somnifera from leaves were established on MS (Murashige and Skoog) media supplemented with Dicamba (2 mg/l), Kinetin (0.1 mg/l) and Sucrose (3% w/v). Suspension cultures were established and the growth and production kinetics were studied. For growth kinetics MS media supplemented with Dicamba (2 mg/l), Kinetin (0.1 mg/l) and Sucrose (3% w/v) without agar was used which was found suitable for the initiation and maintenance of the suspension cultures cultures from the calli. Half B5 medium supplemented with 2,4- Dichlorophenylacetic acid(2,4-D) (1 mg/l), Kinetin (0.1 mg/l) and Sucrose (5% w/v) was employed. Effect of various precursors and elicitors on suspension cultures was studied. The addition of precursor’s sodium acetate (50 mg/l) (~10 folds), Mevalanolactone (50 mg/l) (~14 folds), Squalene (50 mg/l) in colloidal form (~23 folds) and Cholesterol (25 mg/l) in colloidal form (~30.5 folds) have shown significant increase in bioproduction of withaferin A. Among the various biotic and abiotic elicitors used Verticillium dahliae (5% w/v cells extract) (~10 folds) and copper sulphate (100μm/l) (~2.5 folds) have shown moderate increase in the bioproduction of withaferin A. The effective precursors and elicitors were studied for optimization of day of addition and found that maximum bioproduction of withaferin A was seen in the cultures when precursors and elicitors were added to 3 day old suspension cultures.