Ganoderma sinense, a well-known medicinal macrofungus of Basidiomycetes, is widely used in traditional medicine for promoting health and longevity in East Asia. The fruiting bodies of G. sinense contain polysaccharides, ergosterol, and coumarin, which have antitumor, antioxidant, and anticytopenia activities. Mushroom cultivation requires suitable conditions for the formation of fruiting bodies and yield. However, little is known about the optimal culture conditions for mycelial growth and cultivation of G. sinense. In this study, the successful cultivation of a G. sinense strain collected from the wild was reported. The optimal culture conditions were identified by examining one factor at a time. The results of this study revealed that the nutritional requirements for the optimal mycelial growth of G. sinense were fructose (15 g/l) as the carbon source and yeast extract (1 g/l) as the nitrogen source. The optimal pH and temperature for G. sinense were 7 and 25-30EC, respectively. The mycelia grew fastest in treatment II (69% rice grains + 30% sawdust + 1% calcium carbonate). G. sinense produced fruiting bodies under all tested conditions and showed the highest biological efficiency (2.95%) in treatment B (96% sawdust, 1% wheat bran, 1% lime). In summary, under optimal culture conditions, G. sinense strain GA21 showed satisfactory yield and a high potential for commercial cultivation.