Gymnema sylvestre R.Br. (syn. Periploca sylvestris Retz), sometimes referred to as Gudmar, is a main medicinal plant utilised under various medical systems as a treatment for the curing of diabetes mellitus. The culture of selected explants namely shoot apex, nodal stem, leaf and inflorescence on five selected media having MS basal salts supplement with several concentrations of cytokinins (KIN and BAP) plant growth regulators, effected on the establishment of aseptic culture, swelling of explants, disorganized proliferation of cells from the cultured explants and shoot bud growth from the shoot apex and nodal stem explants. The best establishment of aseptic culture was observed on medium M5 (MS + 1.5 “mgl-1” BAP + 0.5 “mgl-1” KIN). The highest swelling of the cultured explants was observed on medium M4 (MS + 2.0 “mgl-1” BAP + 1.0 “mgl-1” KIN). The best caulogenesis was observed on medium M3 (MS + 2.0 “mgl-1” BAP + 0.5 “mgl-1” KIN). Cultured explants leaf were the best for establishment of cultures, swelling and caulogenesis induction. The development of existing shoot bud was the maximum in medium M5 and shoot apex showed better shoot growth compared to nodal stem. The developed shoots can be used as source of new explants and their culture will result in propagule multiplication. Finally, the in vitro developed shoots can be rooted leading to the establishment of micropropagation method in this important remedial plant.