Withaniasomnifera (L.) Dunal. (Indian ginseng) is one of the most important medicinal plants used as a crude drug for its preventive and therapeutic purposes. Among the diverse constituents of Withania, withanolides are found to be the major components responsible for their biological and pharmacological actions. The present study deals with the effect of inoculum density and different media on the growth of hairy roots and withanolide-A production from Withaniasomnifera. An inoculum size of 10 g/L FW favoured the biomass accumulation(120.42 g/L of FW and 11.98 g/L DW) and withanolide-A production(11.96 mg/g DW) inthe tested range of 2.5, 5.0, 10.0 and 20.0 g/L FW. Among different media tested [Murashige and Skoog (MS), Gamborg’s (B5), Nitsch and Nitsch (NN) and Chu’s (N6)], MS medium favoured both biomass accumulation (121.15 g/L FW and 11.96 g/L DW) and withanolide-A production (11.50 mg/g DW).