Plasmodium falciparum Parasite Cultivation

All experiments were performed using Plasmodium falciparum Dd2 (Wellems et al., 1990 (link)), ACP_L-GFP D10 (Waller et al., 2000 (link)), or ACP_L-GFP PfMev NF54 (Swift et al., 2020 (link)) parasite strains, which were obtained from colleagues and verified by confirming their expected drug sensitivity and/or sequencing strain-specific genetic markers. Parasite culturing was performed as previously described (Sigala et al., 2015 (link)) in Roswell Park Memorial Institute medium (RPMI-1640, Thermo Fisher 23400021) supplemented with 2.5 g/L Albumax I Lipid-Rich BSA (Thermo Fisher 11020039), 15 mg/L hypoxanthine (Sigma H9636), 110 mg/L sodium pyruvate (Sigma P5280), 1.19 g/L HEPES (Sigma H4034), 2.52 g/L sodium bicarbonate (Sigma S5761), 2 g/L glucose (Sigma G7021), and 10 mg/L gentamicin (Invitrogen Life Technologies 15750060). Cultures were maintained at 2% hematocrit in human erythrocytes obtained from the University of Utah Hospital blood bank, at 37°C, and at 5% O₂, 5% CO₂, 90% N₂. Cultures were mycoplasma-free by PCR test.