Abstract: A protocol has been developed for in vitro shoot proliferation from callus cultures of Podophyllum hexandrum Royle. Callus initiation occurs from root segments of established in vitro grown seedlings on Gamborg`s B5 media (half strength) supplemented with 2,4-D (0.5-1.5 mg L-1) and BA (0.2-1.0 mg L-1). The rhizome of this plant contains several important lignans and most important being podophyllotoxin, main precursor for anticancer drugs teniposide and ertoposide. In addition Podophyllum hexandrum has been reported to have radioprotection properties. Ruthless collection has led to the disappearance of this important medicinal plant from many areas of Himalayas. Thus there is need for immediate conservation of this important medicinal plant through tissue culture means. In the present study shoot proliferation occurs from callus cultures cultured on basal MS medium fortified with BA and IAA either alone or in combination (0.5-5.0 mg L-1) each. Regenerated shoots were rooted with high efficiency on MS medium fortified with activated charcoal (0.5-1.0%) and NAA (0.5-2.0 mg L-1). The rooted plantlets were transferred to green house in jiffy pots containing sand, soil and vermiculite in 1:1:1 ratio.