Volvariella bombycinais an edible mushroom commercially cultivated in Korea and other countries. In this study, the culture characteristics and optimal growth conditions of milky mushroom strains were determined. The growth temperature and pH range of V. bombycinawas extensively investigated between 15 and 35oC and pH 3–11. For efficient cultivation, 20 types of nutrient sources were selected, specifically, 21 carbon sources, 6 organic nitrogen sources, 7 inorganic nitrogen sources, 13 amino acids, 9 organic acids, and 13 inorganic salts. The impact of each of the selected nutrition sources and their concentration on growth was investigated. The optimum temperature and pH were determined to be 30oC and pH 8.0, respectively. The optimum concentration of medium elements required for mycelial growth of V. bombycinawas determined to be as follows: carbon source, 0.7% Soluble starch; organic nitrogen source, 0.7% Soytone; inorganic nitrogen source, 1.0% (NH4)2C4H4O6; amino acid, 0.1% cysteine; organic acid, 0.07% lactic acid; and inorganic salt, 0.07 mM CaCl2.