Present studies are carried out to find media components and culture methods for in vitro propagation of Athyrium niponicum and to establish the optimal economic masspropagation systems. Among pinnae, petiole and rhizome segments only rhizome segments produced young plants. Rhizome segments showed vigorous plant regeneration on 1/2MS medium and supplement to 1% sucrose and 50 $mg{cdot}L^{-1}$ $NaH_2PO_4$ were promoted the plant regeneration from rhizome segments. Kinetin was better than BA for plant regeneration and combination with 2 ${mu}M$ kinetin and 5 ${mu}M$ IBA was most efficient for plant regeneration. Solid or liquid medium with or without 0.1% qactivated charcoal in modified 1/2MS medium (1% sucrose, 50 $mg{cdot}L^{-1}$ $NaH_2PO_4$, 2 ${mu}M$ kinetin, 5 ${mu}M$ IBA, pH 5.8) were used to find the optimal culture methods. The plant regeneration from rhizome segments were most vigorous on solid medium without activated charcoal. The addition of activated charcoal were inhibited the plant regeneration from rhizome segments not only on solid medium but also liquid stationary or suspension culture.