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00925njm 22002777a 4500 dc Radfar, Azar author Méndez, Dario author Moneriz, Carlos author Linares Gómez, María author Marín-García, Patricia author Puyet Catalina, Antonio author Díez Martín, Amalia author Bautista Santa Cruz, José Manuel author 2009 This protocol describes a method for preparing cultures of Plasmodium falciparum synchronized at any intraerythrocytic stage. Using this method, around 60% parasitized cells may be obtained. On the basis of Trager and Jensen's original continuous culture method, our approach relies on the use of fresh human blood not older than 2 weeks, a low hematocrit between 0.8 and 1.5%, a starting frozen inoculum of 10% ring-stage parasitemia, human serum replaced with AlbuMAX I and alternating sorbitol and Percoll synchronization methods to shorten the cycle window to 4–6 h and reduce sorbitol toxicity. From our synchronized high parasite density cultures, 3–5 ml of infected red blood cells can be obtained in 1 week, corresponding to 1.2 mg of total parasite protein per ml of harvested culture. On the basis of the variables parasitemia and packed cell volume, we provide an equation to accurately calculate the amount of complete medium required every 24 h corrected for the cycle stage and capacity of the culture flask. Ten days suffice to complete the protocol from a frozen stock of parasites. Radfar, A., Méndez, D., Moneriz, C. et al. Synchronous culture of Plasmodium falciparum at high parasitemia levels. Nat Protoc 4, 1899–1915 (2009). https://doi.org/10.1038/nprot.2009.198 1754-2189 10.1038/nprot.2009.198 https://hdl.handle.net/20.500.14352/96172 1750-2799 https://doi.org/10.1038/nprot.2009.198 20010926 Synchronous culture of Plasmodium falciparum at high parasitemia levels