Abstract
In vitro culture of asexual blood stages of Plasmodium falciparum is essential to study the parasite biology, and several aspects need to be addressed to successfully cultivate the parasites, including the requirements for red blood cells and specific nutrients. Since Trager and Jensen established the technique in 1976, some modifications have been introduced to improve the growth rate and yield. In brief, the method is based on the use of human red blood cells suspended in RPMI-1640 culture medium supplemented with a source of lipids and maintained at 37 °C in a low-oxygen atmosphere. In this protocol, a description of thawing, culturing, and cryopreservation of asexual blood stages of P. falciparum is presented.
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Acknowledgments
The author’s research work has been supported by the Danish International Development Agency—Danida (17-02-KU), and Lundbeck Foundation (R250-2017-1289). We thank Maiken Visti for her technical assistance; Michael Ofori (Noguchi Memorial Institute for Medical Research, University of Ghana) for blood collection; Asier García and Anne Knudsen (Faculty of Health and Medical Sciences, University of Copenhagen) for comments and suggestions.
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Lopez-Perez, M., Seidu, Z. (2022). Establishing and Maintaining In Vitro Cultures of Asexual Blood Stages of Plasmodium falciparum. In: Jensen, A.T.R., Hviid, L. (eds) Malaria Immunology. Methods in Molecular Biology, vol 2470. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2189-9_5
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DOI: https://doi.org/10.1007/978-1-0716-2189-9_5
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