Different experiments were conducted to test the effect of growth regulators with the different concentration and combination of (0.5-2.0 mg/l) 2, 4-D with cytokinins like 0.5 mg/l BAP and 0.5 mg/l KN on indirect organogenesis response from leaf segments of Spilanthes acmella. On the most media combinations, leaf segments either callused or differentiated into shoots. Maximum callus (70.5%) was induced on the media containing 1.5 mg/l 2, 4-D with 0.5 mg/l BAP within 30 days and the callus formed was dark green and fragile. After the callus formation the induced callus were sectioned into small pieces and transferred on MS media supplemented with BAP and KN separately or BAP and KN with NAA, IBA and IAA. The best response (75.0%) on shoot regeneration from callus was observed on MS medium supplemented with 1.5 mg/l BAP and 0.5 mg/l IBA with the formation of 6.5±0.41 number of shoots having length of 7.9±0.1 centimeters within 20 days. after shoot regeneration maximum (75.0%) of shoot multiplication rate was observed with an average of 7.5±0.41 adventitious shoots, directly from the explants, without any callus formation on medium supplemented with 1.0 mg/l BAP and 0.5 mg/l IBA. Shoots formed were remarkable healthy and achieved the length of 8.5±0.1centimetres within 20 days. While as maximum (70.0%) root induction was achieved directly from the base of the regenerated shoots on medium supplemented with 0.5 mg/l IBA of average length 6.0±0.6 centimetres within 20 days.