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Plasmodium falciparum: Modifications of the In vitro Culture Conditions Improving Parasitic Yields

J. Werner Zolg, Alexander J. MacLeod, Ida H. Dickson and John G. Scaife
The Journal of Parasitology
Vol. 68, No. 6 (Dec., 1982), pp. 1072-1080
DOI: 10.2307/3281094
Stable URL: http://www.jstor.org/stable/3281094
Page Count: 9
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Plasmodium falciparum: Modifications of the In vitro Culture Conditions Improving Parasitic Yields
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Abstract

We attempted to optimize some of the variables involved in the in vitro culturing of Plasmodium falciparum. Irrespective of the isolates used, suspension cultures in glucose-enriched RPMI-1640 medium buffered with TES yielded about twice the amount of parasites than could be obtained from static, thin-layer cultures with HEPES-buffered RPMI-1640 without additional glucose. In suspension cultures, methylcellulose (1 mg/ml) as added to protect the erythrocytes. In addition the erythrocytes were found to be more suitable for culturing P. falciparum when stored as a concentrate in saline-adenine-glucose than as whole blood in citrate-phosphate-dextrose. With a cloned isolate of P. falciparum (Tak9/clone 96) a further stimulation of the final parasitemia could be achieved by supplementing the medium with hypoxanthine (50 µg/ml) and reduced glutathione (600 µg/ml). Moreover, we identified hypoxanthine and glutathione as two of the factors critical for the ability of human serum to support the growth of the parasites. These modifications give a two- to four-fold increase in the final parasitemia over the original Trager-Jensen culture method, thus allowing more parasites to be isolated for biochemical studies.

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