1 Muhimbili Sickle Cell Programme, Muhimbili University of Health and Allied Sciences, Tanzania.
2 Department of Biochemistry, Muhimbili University of Health and Allied Sciences, Tanzania.
3 Department of Parasitology, Muhimbili National Hospital.
4 Department of Parasitology and Medical Entomology, Muhimbili University of Health and Allied Sciences, Tanzania.
5 Department of Hematology and Blood Transfusion, Muhimbili University of Health and Allied Sciences, Tanzania.
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Malaria morbidity and mortality, almost entirely from Plasmodium
falciparum, are still rampant in Africa: therefore, it is important to
study the biology of the parasite and the parasite-host cell
interactions. In vitro cultivation of Plasmodium falciparum is most
useful for this purpose, as well as for investigating drug resistance
and possible new therapies. Here we report that the Trager & Jensen
continuous culture of P. falciparum can be established in a laboratory
in Tanzania with minimal facilities and with modest expenditure.
|Figure 1. Selection of Clinical malaria samples for in vitro cultures.
|Figure 2. Laboratory set-up for Plasmodium falciparum culture. A: Telstar Biosafety Cabinet Class II A with UV light on when not in use. B: Same cabinet when in use. C: Close-up of candle jar with lighted candle. D: Candle jar (flame off) in 37 °C incubator.|
1. List of sources for Plasmodium falciparum cultures.
|Table 2. Data on individual culture attempts from clinical samples in year 2020.|
|Figure 3. Microscopic images of Plasmodium falciparum from in vitro cultures A: Low power view of culture smear: all stages of parasite development are seen. B: High power view: red cell with multiple rings C: Schizont with hemozoin (malarial pigment).|
|Figure 4. Exponential growth of Plasmodium falciparum in cultures from clinical isolates. A mixture of serum/albumax 50:50 was used for culturing. Values on the ordinate axis were calculated from parasite counts on smears, taking into account the dilution of parasitized red cells by non-paraistized red cells each time fresh red cells were added every 2 days.|
|Figure 5. Supplementation of media with a mixture of human serum and bovine serum optimizes parasite growth. Parasites were cultured in complete culture medium containing i) Albumax II ii) 50:50 Albumax II and human serum A+ and iii) Human Serum A+. The experiment was performed in two replicates. Paired t-test was used to compare the mean differences. Serum vs Albumax (p=0.0567); 50:50 vs serum (p=0.0123) and 50:50 vs Albumax (0.0636).|
1. Growth of Plasmodium falciparum
in cultures from clinical isolates. The actual parasitemia is presented
with days in which the culture was diluted with uninfected red blood
cells (presented in bold and italics).