Abstract: Propagation of Sauropus androgynus (SA) by tissue culture technology provides a better alternative than conventional propagation, as it ensures uniformity in production of sustainable plantlets with desirable traits. The objective was to establish an efficient plant regeneration protocol using leaf, nodal and internodal explants of SA, starting from the establishment of aseptic cultures to the acclimatization stage of plantlets. The effects of plant growth regulators on the growth performance of SA were studied. Results showed in vitro mass multiplication of SA was achieved via indirect organogenesis and shoot regeneration pathway. After 30 days of culture inoculation, high percentage of aseptic cultures were established from nodal (85%), internodal (85%) and leaf (91%) explants sterilized with 70% (v/v) ethanol for 1 min followed by 20% (v/v) Clorox for 20 min. MS medium supplemented with 2.0 mg L–1 BAP (6-benzylaminopurine) and 0.5 mg L–1 IAA (indole-3-acetic acid) proved to be effective in inducing adventitious shoots from internodal (95%), nodal (96%) and leaf (90%) explants, after 60 days of culture. High frequency of callus formation (38%) was obtained from leaf explants cultured on MS medium containing 2.0 mg L–1 IAA and 0.5 mg L–1 BAP. Rooting response of 90% was achieved from cultured shoots using half-strength MS medium enriched with 1.0 mg L–1 IAA and from these, 85% survived acclimatization and grew vigorously after 4 weeks. This optimized protocol established a complete in vitro plant regeneration of SA and has never been reported previously. This may form the basis for future research in germplasm conservation and genetic transformation for sustainable production of high quality SA.