Mycelial-form Sporothrix schenckii was studied to determine if growth in complex (Sabouraud-dextrose) or defined culture media (minimal medium, Gibco Medium 199 with and without Hepes buffer) with differing initial pH levels would affect expression of antigen components. Cultures were evaluated by continuous monitoring and serial sampling for various parameters. Great variation was seen in the protein and antigenic patterns induced by the different media. The expression of a 55 kDa component accompanied by significant acidification of the culture medium at the beginning of the stationary growth phase was notable in Sabouraud medium. In the chemically defined media, a 90 kDa component was expressed that reacted with sera from patients with sporotrichosis. The pH in these media showed little change during the different growth phases of the fungus. Among the media studied, minimal medium favored the expression of the greatest number of antigenic components. In all of the assays, the stationary growth phase appeared to be optimal for content of antigenic components. Cross reactions were not observed with any of the culture filtrates using sera from patients with other mycoses.