Use of Gymnema sylvestre, commonly known as Periploca of woods an Indian medicinal woody climber has increased recently due to the pharmaceutical potential of gymnemic acids, found in its leaves. Gymnemic acids has been reported to effect a natural treatment for diabetes. This study developed a novel cell culture system for in vitro growth and production of this species, suggesting a possible technology for large scale production of gymnemic acids. Leaf explants grown in Muragish and Skoog salts supplemented with IAA 1.5 mg L-1 and BA 0.5 mg L-1 gave maximum percentage callus formation compared to other medium treatments evaluated. Combination of external phytohormone, shaking speeds, pH of the medium, played important roles in growth and gymnemic acid production in suspension cell cultures. © 2006 Asian Network for Scientific Information.