The Parasitology Core Facility (PCF) offers infectious Plasmodium falciparum gametocyte cultures that can be used for infection assays in mosquitoes.
We are using Plasmodium falciparum NF54 strains and generate gametocyte cultures that contain 2-4% mature stage V gametocytes. Orders for gametocyte cultures have to be placed 3 weeks in advance since 15 days are required for mature gametocytes to form. A culture request/order provides 25-30ml of cultured parasites. Gametocyte cultures are initiated from asexual cultures by a process known as bulking to induce formation of gametocytes. Bulking simply means that the volume of asexual culture media is increased without adjusting the hematocrit concentration. Media is changed daily until gametocytes mature (15 days minimum). Cultures are kept at 37°C and gassed using the standard candle jar method. When gametocytes are ready, the percentage gametocytemia is determined by counting mature (stage V) gametocytes in fields containing at least 500 rbc. To prepare the culture for mosquito infection, gametocytes are adjusted to 0.3% and feeds are done at a final hematocrit concentration of 50%. The rbc and normal human serum for mosquito feeding has to be kept at 37°C to prevent premature exflagellation of male gametocytes.
Our cultures will normally generate up to 100 - 300 oocyts per mosquito midgut. However, mosquito infection efficiency is also highly dependent on how the mature culture is handled prior to feeding. Once cultures are delivered, it is the user’s responsibility to ensure that gametocytes are properly handled and diluted (general guidelines above) to ensure optimal mosquito infections. The PCF core personnel offer training on mosquito feeding and can also perform feeds at a nominal fee.
Pricing for gametocyte culture: $100 for one plate 25 ml culture
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