Celastrus paniculatus Willd, a vulnerable important woody liana with high medicinal value and limited reproductive capacity has been over exploited due to its demand in the pharmaceutical industry. In the present study, an efficient micropropagation protocol was developed from nodal explants on Murashige and Skoog and woody plant medium. The effect of various strengths of WP and MS medium (total, half strength) and different concentrations of plant growth regulators including cytokinins [6-benzylaminopurine (BAP) and Kinetin) and auxins (α-naphthaleneacetic acid (NAA) and (IBA) indole-3-butyric acid] on proliferation and rooting was investigated. C. paniculatus shoots showed maximum proliferation in WP medium supplemented with BAP (0.5 mg l-1). Best rooting frequency and higher root number and length were attained in medium fortified with NAA in combination with 0.2% activated charcoal. Seventy percent of the plantlets were successfully acclimatized to ex vitro conditions, exhibiting normal development. These plantlets can be used to replenish the declining populations in the wild. Molecular profiling of the in vitro developed plantlets was done using RAPD to confirm their genetic uniformity.