Optimal culture conditions for efficient in vitro propagation and polysaccharide production of Orostachys japonicus were established. The highest growth yield was achieved in 1/2 MS medium, while the lowest growth yield was obtained in 4 MS medium. The patterns of polysaccharide formation were a little similar in all cases, but on MB5 medium, the po]ysaccharide contents of plant were higher than others. Among the modified nitrate levels, effective growth level were obtained in 1/4 N and 1/2 N. High contents of polysaccharide were obtained in 4 N. Different concentration of potassium and calcium did not improve the growth and polysaccharide production. The micropropagated shoots were successfully acclimatized artificial soils.