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  • Plasmodium falciparum Gametocyte Culture and Mosquito Infection Through Artificial Membrane Feeding.

Plasmodium falciparum Gametocyte Culture and Mosquito Infection Through Artificial Membrane Feeding.

Journal of visualized experiments : JoVE (2020-07-28)
Abhai K Tripathi, Godfree Mlambo, Sachie Kanatani, Photini Sinnis, George Dimopoulos
ABSTRACT

Malaria remains one of the most important public health problems, causing significant morbidity and mortality. Malaria is a mosquito borne disease transmitted through an infectious bite from the female Anopheles mosquito. Malaria control will eventually rely on a multitude of approaches, which includes ways to block transmission to, through and from mosquitoes. To study mosquito stages of malaria parasites in the laboratory, we have optimized a protocol to culture highly infectious Plasmodium falciparum gametocytes, a parasite stage required for transmission from the human host to the mosquito vector. P. falciparum gametocytes mature through five morphologically distinct steps, which takes approximately 1-2 weeks. Gametocyte culture described in this protocol is completed in 15 days and are infectious to mosquitoes from days 15-18. These protocols were developed to maintain a continuous cycle of infection competent gametocytes and to maintain uninterrupted supply of mosquito stages of the parasite. Here, we describe the methodology of gametocyte culture and how to infect mosquitoes with these parasites using glass membrane feeders.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Xanthurenic acid, 96%
Sigma-Aldrich
Hanks′ Balanced Salt solution, Modified, with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
N-Acetyl-D-glucosamine, suitable for cell culture, BioReagent
Sigma-Aldrich
Sodium bicarbonate, BioXtra, 99.5-100.5%