Pure spherules of Coccidioides immitis in continuous culture

Investigation of host-parasite relationships involving the parasitic form of Coccidioides immitis has been difficult because, previously, spherules and endospores have not been grown continuously in tissue culture medium without detectable formation of hyphae. Arthroconidia were harvested from mycelial cultures and inoculated into tissue culture flasks which contained RPMI 1640 medium supplemented with 10% calf serum and N-Tamol (Rohm & Haas Co., Philadelphia, Pa.). Flasks were purged with 5% CO2, sealed, and placed on a reciprocating shaker at 35 degrees C. Hyphae which arose during incubation were removed by filtration. Arthroconidia readily converted to the spherule-endospore form within 12 days. Six days after complete conversion, spherules and endospores were transferred to RPMI 1640 without N-Tamol. The spherule-endospore cycle was maintained in tissue culture medium for 84 days without the formation of detectable hyphae.