Present investigation deals with establishment of standard protocol for high frequency regeneration in medicinal plant Tylophora asthmatica (L. f.) Wight & Arn. Regeneration of plant has achieved on MS-media in combination with various concentrations of BAP and KIN along with IAA. The most suitable concentration was recorded at 3.0 mg/L of BAP along with IAA for high frequency of regeneration. KIN along with IAA facilitates multiple shooting along with somatic embryogenesis. Rhizogenesis were achieved using IAA and NAA and plants hardening protocol was followed and regeneration of plants was achieved.