Currently, the spawn of the mushroom Agarcus bisporus is produced by a method developed in the 1980s, and anew spawning method needs to be developed to improve the quality of the spawn. In this study, the condition for a maximum mycelium weight(5.92±0.52 g/L) was shaking culture (24 hours/day) at 24°C and 120 rpm in CDB (compost dextrose broth). Based on this, the ventilated liquid culture method (2.5 L/min) was cultured for 10 days. This method was appropriate, andwhen the inoculum was cultured at 50 g/mL for about 10 days, it was cultured well without agglomeration and shaking of seed.