Culturing Toxoplasma gondii Tachyzoites

Toxoplasma gondii was maintained by serial passage of tachyzoites in HFF monolayers cultured in EMEM containing 1% FBS, (2 mM glutamine, 100 units/ml penicillin, and 100 μg/ml streptomycin) as previously described [7 (link)]. Uracil auxotrophs were supplemented with 0.2 mM Uracil (Sigma-Aldrich). OMP [13 (link)], CPS [5 (link)], CPS-YFP [96 (link)], and other strains used for treatment of ID8DV tumors were cultured in HFF monolayers and freshly lysed extracellular tachyzoites were purified by filtration through 3.0 μm filters (Nuclepore), and washed extensively in Dulbecco's Phosphate Buffered Saline (PBS) prior to intraperitoneal injection in 0.2 ml PBS in ID8DV ovarian tumor-bearing mice.