RT Journal Article
T1 Synchronous culture of Plasmodium falciparum at high parasitemia levels
A1 Radfar, Azar
A1 Méndez, Dario
A1 Moneriz, Carlos
A1 Linares Gómez, María
A1 Patricia Marín-García, 
A1 Puyet Catalina, Antonio
A1 Díez Martín, Amalia
A1 Bautista Santa Cruz, José Manuel
AB 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.
SN 1754-2189
YR 2009
FD 2009
LK https://hdl.handle.net/20.500.14352/96172
UL https://hdl.handle.net/20.500.14352/96172
LA eng
NO 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
DS Docta Complutense
RD 22 jul 2024