Using nodal segment and leaf segment explant, callus cultures were initiated. They were grown on MS medium containing 3% sucrose supplemented with different concentrations of growth hormones (0.10, 0.25, 0.50, 1.0, 1.50, 2.50, and 5.0 mg/L). Various growth hormones used are 2,4-dichlorophenoxy acetic acid (2,4-D), Naphthalene Acetic Acid (NAA), Indole-3-Acetic Acid (IAA) and 6-benzylaminopurine (BAP). The high biomass yielding concentration was used to observe the effect of plant growth regulators in combination. Among the combinations used, high biomass yield was obtained in 0.5 mg/L 2,4-D + 0.5 mg/L BAP + 1.5 mg/L IAA. Higher amount of gymnemic acid was obtained in 0.5 mg/L 2,4-D + 0.5 mg/L BAP combination. The harvested cell biomass and genuine samples were subjected to the extraction of the active principle. The High Performance Thin Layer Chromatography (HPTLC) analysis showed that the quantity of gymnemic acid is higher in genuine samples than callus.

Herbal medicines are becoming more popular in the developing countries of the world because of their efficacy, safety and lesser side effects, as these medicines has attune with human body. The increasing awareness about side effects of drugs has made the western pharmaceutical industries turn towards the herbal-based Indian and Chinese medicine. The natural habitats for medicinal plants are disappearing at a faster rate because of the environmental and geographical instabilities, mainly due to their overexploitation. Gymnema sylvestre strands are rapidly being depleted because of the high demand for them from pharmaceutical companies. This has prompted the industries as well as scientists to consider the possibility of in vitro techniques as an alternative supply for the fast production of plantlets and raw material for pharmaceuticals. This has created a considerable interest in plant tissue culture as a potential alternative source to traditional agricultural practices. These facts demand alternative ways for their production.

For the large production of secondary metabolites, in vitro callus culture system can be used. G. sylvestre is a highly valuable medicinal plant which belongs to the family Asclepiadaceae. As per the International Union for Conservation of Nature (IUCN), it falls under the Rare Endangered and Threatened (RET) category. In previous literature,G. sylvestre is mentioned as a vulnerable species. It is a slow growing perennial, medicinal woody climber. It is found to be growing in central and peninsular India. A review of the literature reveals that it consists of many medicinal properties because of the presence of triterpene saponins.

Biotechnology Journal, Vitro Production, Callus Biomassby, Plant Growth Regulators, Gymnemic Acid Quantity, Gymnema Sylvestre, Naphthalene Acetic Acid, NAA, Indole-3-Acetic Acid, IAA, Herbal Medicines, Pharmaceutical Industries, Gymnema Sylvestre, International Union for Conservation of Nature, IUCN, Rare Endangered and Threatened, RET, Dichlorophenoxyaceticacid, Gymnemic Acid.