The present investigation deals with in vitro regeneration of an epiphytic orchid, Dendrobium aphyllum (Roxb.) C.E.C Fischer using leaf explants through Protocorm-like bodies (PLBs) proliferation. The species collected from the Darjeeling Himalaya (Latpanchar forest, Kurseong) has been classified as Least Concern by the IUCN Red List (2020). Ethnomedicinally, this plant has been used for curing skin disorders, scorpion bite, and wounds. Additional presence of 2,4-D/BAP (0.5, 1.0, 1.5, 2.0 mgl-1) in MS medium proved obligatory for inducing callusing in the leaf explants; callusing was however, highly promoted in the former combination. Further, the combination of both these additives showed synergistic effect in inducing better response. The growth and differentiation of callus tissues derived from leaf explants was further investigated on MS medium supplemented with different concentrations of 2,4-D and BAP. Optimal results were obtained on 2, 4-D (1.5 mgl-1) and BAP (1 mgl-1) combination; the callus differentiated Protocorm-like bodies (PLBs). After subculturing, calli initially exhibited 3 different appearances: light green, yellow, and light brown. The light green calli grew rapidly, consisting compact mass of isodiametric granules, and produced 1.2 PLBs on an average per 0.5 cm diameter of callus on MS medium without growth regulators. Subsequently, the PLBs were inoculated on 4 different nutrient media such as MS, Gamborg B5, Vacin and Went, and M for the differentiation of shoots and roots. After 6 wks, about 4.2 plantlets were formed from each PLB on MS basal medium; the highest shoot length (7.4 cm) and a greater number of leaves (3.8) on average were observed in the plantlets in this combination. The regenerated plantlets displayed maximum survival frequency (100%) in the potting medium with the combination of charcoal and brick pieces. The method can be successfully used for mass propagation of the species.