This study was carried out to obtain basic data on physiological characteristics for an artificial cultivation of Grifola frondosa. Ten strains of G. frondosa were collected from Korea, China and Japan and those were isolated and tested. PDA medium was selected for the favorable culture medium. The optimal conditions for the mycelial growth of G. frondosa were 25℃ and pH 4∼5 respectively. The carbon and nitrogen sources for the optimum mycelial growth were fructose and peptone, respectively. Highest mycelial growth was observed when the C/N ratio was 10∼20. Significant differences among lines were found for production yield and crop cycle time. KME44009 had a short spawn run time, high yield, good color and quality compare with other strains. To find out the suitable medium composition of G. frondosa, physicochemical conditions were investigated. The T2 formulation (55:25:8:12 mixing ratio of oak sawdust, oak chip, wheat bran and soybean cake) showed the shortest time to complete the crop cycle and the highest yields(weight of fresh mushrooms harvested at maturity). Among the physicochemical factors, pH, crude oil and total nitrogen may affect mushroom yield. Therefore, development of suitable medium would benefit from increased production efficiency of G. frondosa mushrooms and be commercial potential.