Gametocytogenesis of Plasmodium falciparum in vitro: a simple technique for the routine culture of pure capacitated gametocytes en masse

Abstract

SUMMARYA technique is described for the growth of pure gametocyte cultures of Plasmodium falciparum. Using the classification of Hawking, Wilson & Gammage (1971) these cultures contain gametocytes of stages III, IV and V alone. Routine sexual cultures, varying from 0·35 ml static cultures to 500 ml shaking cultures, are exposed to an inhibitor of DNA synthesis (mitomycin C at 10 μg/ml) on the 11th day of culture, and the culture is harvested on the 14th day when capacitated stage V gametocytes are present. All other stages are killed by the drug and are morphologically degenerate within 2 days of the addition of the inhibitor.

The cell biology of malaria infection of mosquito: advances and opportunities

“…Have we identified all the key regulator(s) of sexual differentiation, certainly not, but we are now developing the tools for doing so. Despite long-established protocols for the culture of P. falciparum gametocytes en masse (Ponnudurai et al, 1983;Sinden et al, 1984) and the detailed descriptions of the complex morphological changes occurring in the skeletal, cytoplasmic and nuclear organization of the parasites (Sinden et al, 1978;Dixon et al, 2012;Hliscs et al, 2015) and in the organization of the mitochondrion and apicoplast (Okamoto et al, 2008), we remain disturbingly ignorant of the molecular profile of the maturing gametocyte. Transcriptomic and proteomic studies (Hall et al, 2005;Khan et al, 2005;Tao et al, 2014) have revealed, for example, 174 male-and 258 female-enriched proteins in mature (dimorphic/separable) P. falciparum gametocyte preparations.…”