Plasmodium falciparum

Niu, Guodong, Wang, Xiaohong & Li, Jun, 2024, Leucinostatins target PlaSModIUM mitochondria to block malaria transmission, Parasites & Vectors (524) 17 (1), pp. 1-9 : 2

P. falciparum culture and standard membrane feeding assays (SMFA)

P. falciparum (NF54) was cultured in the lab [ 7, 8, 17] with 4% O + - type red blood cells in a complete Roswell Park Memorial Institute (RPMI) 1640 medium with 10% human AB + serum and 12.5 μg/ml of hypoxanthine in a candle jar at 37 °C. Ten, 15-day-old cultured P. falciparum -infected cells were collected through centrifugation (300 g for 3 min), and about 100 μl of pellet were resuspended in 1.5 ml fresh AB + - type serum and mixed with 1.4 ml O + - type red blood cells, containing about 0.2% stage V gametocytes [ 16]. Each candidate compound was dissolved in dimethylsulfoxide (DMSO) and diluted with DMSO to obtain different concentrations. About 2 μl of candidate solution was mixed with 298 μl of P. falciparum -infected blood prepared above and was used to feed 60–80 3–5-day-old An. gambiae with feeders covered with parafilm at 37 °C for 30 min. Te fed mosquitoes were maintained in an insectary with 10% sugar in water for 7 days in the insectary and dissected to obtain midguts. Te midguts were stained with 0.1% mercury dibromofluorescein disodium salt in phosphate-buffered saline (PBS), and the oocysts were counted under a light microscope.