Sorbitol

Asexual blood stage 3D7 P. falciparum parasites were cultured as previously described in (Lopez‐Rubio et al, 2009). Briefly, parasites were cultured in human RBCs (obtained from the Etablissement Français du Sang with approval number HS 2016‐24803) in RPMI‐1640 culture medium (Thermo Fisher 11875) supplemented with 10% v/v Albumax I (Thermo Fisher 11020), hypoxanthine (0.1 mM final concentration, C.C.Pro Z‐41‐M) and 10 mg gentamicin (Sigma G1397) at 4% hematocrit and under 5% O₂, 3% CO₂ at 37 °C. Parasite development was monitored by Giemsa staining. Parasites were synchronized by sorbitol (5%, Sigma S6021) lysis at ring stage, plasmagel (Plasmion, Fresenius Kabi) enrichment of late stages 24 h later, and an additional sorbitol lysis 6 h after plasmagel enrichment. The 0 h time point was considered to be 3 h after plasmagel enrichment. Parasites were harvested at 1–5% parasitemia.

Asexual blood-stage 3D7 P. falciparum parasites were cultured as previously described in Lopez-Rubio et al (link)and W.H.O. (2021) . Parasites were cultured in human RBCs (obtained from the Etablissement Français du Sang with approval number HS 2016-24803) in RPMI-1640 culture medium (11875; Thermo Fisher Scientific) supplemented with 10% vol/vol Albumax I (11020; Thermo Fisher Scientific), hypoxanthine (0.1 mM final concentration, C.C.Pro Z-41-M), and 10 mg gentamicin (G1397; Sigma-Aldrich) at 4% hematocrit and under 5% O₂ and 3% CO₂ at 37°C. Parasite development was monitored by Giemsa staining. Parasites were synchronized by sorbitol (5%, S6021; Sigma-Aldrich) lysis at the ring stage, plasmagel (Plasmion; Fresenius Kabi) enrichment of late stages 24 h later, and an additional sorbitol lysis 6 h after plasmagel enrichment. Parasites were cultured under static conditions with the exception of shaking during the late schizont until an early ring stage. Parasites were harvested at 1–5% parasitemia.