Axenic in vitro cultivation of Trypanosoma vivax trypomastigote forms

Cultures of Trypanosoma vivax stocks were initiated with tsetse-derived metacyclics and with organisms isolated from mouse blood. They were propagated as trypomastigote forms at 35 degrees C initially in the presence of bovine endothelial cells. All stocks were subsequently adapted to axenic culture conditions. The standard medium consisted of HEPES-buffered MEM with 1% MEM non-essential amino acids, containing 2 mM L-glutamine, 1 mM sodium pyruvate, 100 IU/ml penicillin, 100 micrograms/ml streptomycin, 0.2 mM hypoxanthine, 0.01 mM bathocuproine sulphonate, 0.1 mM L-cysteine, 0.1 mM 2-mercaptoethanol and 20% caprine or bovine serum. Cultured trypanosomes were morphologically similar to bloodstream forms found in vivo and they retained infectivity after in vitro cultivation.