Long-term in vitro cultures of blood-stage parasites are so far feasible only for Plasmodium falciparum and P. knowlesi. In this chapter, we describe short-term ex vivo culturing of P. cynomolgi and P. vivax. We also describe long-term in vitro culturing of P. knowlesi as well as some techniques for synchronizing parasites. Cultured parasites can be used for a variety of purposes, e.g., for in vitro drug assays and antibody-mediated growth inhibition assays.
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
The authors wish to thank Hasting Ozwara Suba for setting up the long-term in vitro culture of P. knowlesi, Joyce Verburgh for help in the in vitro drug assays, Els Klooster for providing parasite pictures, and Henk van Westbroek for designing the figures.
Kocken CH et al (2006) Plasmodium vivax: in vitro susceptibility of blood stages to synthetic trioxolane compounds and the diamidine DB75. Exp Parasitol 113:197–200PubMedCrossRefGoogle Scholar
Kosaisavee V et al (2006) Plasmodium vivax: isotopic, PicoGreen, and microscopic assays for measuring chloroquine sensitivity in fresh and cryopreserved isolates. Exp Parasitol 114:34–39PubMedCrossRefGoogle Scholar
Wengelnik K et al (2002) A class of potent antimalarials and their specific accumulation in infected erythrocytes. Science 295:1311–1314PubMedCrossRefGoogle Scholar
Kennedy MC et al (2002) In vitro studies with recombinant Plasmodium falciparum apical membrane antigen 1 (AMA1): production and activity of an AMA1 vaccine and generation of a multiallelic response. Infect Immun 70:6948–6960PubMedCrossRefGoogle Scholar
Mahdi Abdel Hamid M et al (2011) Vaccination with Plasmodium knowlesi AMA1 formulated in the novel adjuvant co-vaccine HT protects against blood-stage challenge in rhesus macaques. PLoS One 6:e20547PubMedCrossRefGoogle Scholar
Kocken CH et al (2002) Plasmodium knowlesi provides a rapid in vitro and in vivo transfection system that enables double-crossover gene knockout studies. Infect Immun 70:655–660PubMedCrossRefGoogle Scholar
Rowe AW et al (1968) Liquid nitrogen preservation of red blood cells for transfusion: a low glycerol-rapid freeze procedure. Cryobiology 5:119–128PubMedCrossRefGoogle Scholar
Braun-Breton C et al (1988) Induction of the proteolytic activity of a membrane protein in Plasmodium falciparum by phosphatidyl inositol-specific phospholipase C. Nature 332:457–459PubMedCrossRefGoogle Scholar