Shoot cultures and callus cultures from roots and leaves of Hemidesmus indicus R. Br. (Asclepiadaceae) were established on Murashige and Skoog medium with various hormonal combinations. The production of antioxidants (lupeol, vanillin, and rutin) in shoot cultures, callus cultures derived from leaf cells and root cells, was compared with root and aerial portions of the parent plant. Shoot cultures and leaf callus cultures produced more antioxidants than root callus cultures. In vitro culture of this species might offer an alternative method for production of these important pharmaceuticals, which would reduce the collection pressure on this rare plant.