This study was carried out to determine the effect of medium solidity, salt strength, sugar and nitrogen sources, and pH levels on in vitro multiplication of pathogen-free yam (Dioscorea opposita Thunb.). Liquid medium was more effective in the growth of plant height, fresh weight, and formation of microbulb than the solid medium. Optimal condition for plant fresh weight, growth, and multiplication axillary bud was in 1MS salt strength with 60 $g{cdot}L^{-1}$ sucrose and half strength of $KNO_3$. Optimal condition for microbulb formation was $frac{1}{2}$ MS salt strength supplemented with glucose 60 $g{cdot}L^{-1}$ and half strength of $KNO_3$. The number of leaves and nodes were sharply increased from 2 to 5 weeks, whereas plant fresh weight was steadily increased from 3 to 11 weeks after inoculation. Microbulbs were formed at 2 weeks after inoculation and continuously increased until 12 weeks.