The purpose of this study is in vitro regeneration of Tylophora indica from leaf explants collected from two year old healthy plant and it place on MS medium with BAP (1 mg/l) and NAA (2 mg/l) induce shoots and roots. MS media combination with BAP (0.5 mg/l) and 2, 4- D (2 mg/l) explant show high callus induction. Root initiation observed BAP (2 mg/l) and 2, 4- D (2.5 mg/l).