Improved synchronous production of Plasmodium falciparum gametocytes in vitro.
The sexual stages of the Plasmodium falciparum life cycle are attractive targets for vaccines and transmission blocking drugs. Difficulties in culturing and obtaining large amounts of sexual stage P. falciparum parasites, particularly early stages, have often limited research progress in this area. We present a new protocol which simplifies the process of stimulating gametocytogenesis leading to improved synchronous gametocyte production. This new method can be adapted to enrich for early stage gametocytes (I and II) with a higher degree of purity than has previously been achieved, using MACS magnetic affinity columns. The protocol described lends itself to large scale culturing and harvesting of synchronous parasites suitable for biochemical assays, northern blots, flow cytometry, microarrays and proteomic analysis.
Fivelman
Quinton L
QL
Department of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, London WC1E 7HT, UK. quinton.fivelman@lshtm.ac.uk
McRobert
Louisa
L
Sharp
Sarah
S
Taylor
Cathy J
CJ
Saeed
Maha
M
Swales
Claire A
CA
Sutherland
Colin J
CJ
Baker
David A
DA
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D008018:Life Cycle Stages; D010963:Plasmodium falciparum / Q000254:growth & development* / Q000302:isolation & purification*