Traditional propagation of Tylophora indica is challenging due to low seed viability, poor germination and difficulty in rooting vegetative cuttings. Micropropagation offers the best solution for rapid and true-to-type plant multiplication. The study aims to standardize a micropropagation protocol in Tylophora indica (Burm. F.) Merrill through indirect organogenesis from leaf explant. This protocol includes sterilization of leaf explant, callus production, shoot regeneration, root initiation and acclimatization. Surface sterilization treatment using 70 % ethanol for 2 min, 0.1 % bavistin for 15 min, 1000 ppm streptomycin for 10 min and 0.1 % HgCl2 for 3 min found the most effective. The best callus, which was dark green, friable and heaviest (3.53 g) was observed in MS media supplemented with 1 mg L-1 Benzyl Adenine (BA). MS media supplemented with 2 mg L-1 BA showed the highest shoot regeneration (65.94 %) and maximum number of shoots per callus (12.90). In vitro regenerated plantlets were inoculated into MS media supplemented with different concentrations of Indole-3-acetic acid (IAA). MS media supplemented with 0.5 mg L-1 IAA was found to be the most effective for rooting (83.33 %) and with the longest roots (7.60 cm). In vitro rooted plantlets were acclimatized successfully after primary and secondary hardening. Primary hardening mixture, which was a mixture of cocopeat and soil in a 1:2 ratio, resulted in 80 % survival rate. After secondary hardening, the overall plant survival rate was 75 %.