Abstract: The experiment was conducted at Plant Biotechnology Laboratoty of Institute of Biological Sciences, Rajshahi University, Bangladesh. Subsequent regeneration performance of two medicinal plants (Aristolochia indica Linn and Hemidesmus indicus R.B were studied throughout the experimental period. Different concentrations and combinations of growth regulators were used in MS medium to observe the callus induction, Callus regeneration and root induction. Among the different concentrations and combinations of growth regulators, the highest percentage of callus induction was 90.00% on MS medium supplemented with 1.0 mg L-1 NAA and 1.0 mg L-1 BAP from Aristolochia indica where as 95.00% on MS medium supplemented with 0.5 mg L-1 NAA and 2.0 mg L-1 Kn from Hemidesmus indicus. The highest percentage (85.00) of shoot regeneration was obtained in MS medium fortified with 1.0 mg L-1 BAP and 2.5 mg L-1 NAA from Aristolochia indica where as 95.00% on MS medium supplemented with 2.5 mg L-1 Kn and 1.0 mg L-1 NAA from Hemidesmus indicus. Regenerated shoots rooted best on MS medium containing 2.5 mg L-1 Kn and 2.0 mg L-1 IBA. Plantlets were transferred to pots containing sand and soil mixture, acclimatized in a culture room and finally rooted plants were transferred to soil.