Malaria pp 27-33 | Cite as

In Vitro Differentiation of Plasmodium falciparum Gametocytes into Ookinetes

  • Anil K. Ghosh
  • Marcelo Jacobs-LorenaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 923)


The ookinete is the motile form of the malaria parasite that invades the mosquito midgut epithelium to initiate sporogony. Differentiation of ingested gametocytes into ookinetes in the mosquito midgut lumen and the subsequent interaction with the luminal surface of the midgut epithelium in preparation for invasion are complex processes. To facilitate the study of these events in detail, it is necessary to produce sufficient numbers of pure, fully mature ookinetes. However, production of even a small number of Plasmodium falciparum ookinetes in vitro has proven to be a daunting task. Consequently, over the past four decades our collective understanding of the biology of this parasite form remains sorely deficient. Here, we describe a new culture technique, which improves the in vitro transformation efficiency of P. falciparum gametocytes into mature ookinetes and supports the complete development of ookinetes that retain the ability to infect the mosquito midgut and to produce oocysts.

Key words

P. falciparum gametocytes Ookinetes Gametocytes to ookinete transformation 



This work was supported by grants from the National Institutes of Health, by the Johns Hopkins Malaria Research Institute and by the Bloomberg Family Foundation.


  1. 1.
    Ghosh A et al (2000) The journey of malaria parasite in the mosquito: hopes for the new century. Parasitol Today 16:196–201PubMedCrossRefGoogle Scholar
  2. 2.
    Janse CJ et al (1986) Rapid repeated DNA replication during microgametogenesis and DNA synthesis in young zygotes of Plasmodium berghei. Trans R Soc Trop Med Hyg 80:154–157PubMedCrossRefGoogle Scholar
  3. 3.
    Carter EH et al (1987) The in vitro cultivation of P. falciparum ookinetes, and their enrichment on Nycodenz density gradients. Parasitology 95:25–30PubMedCrossRefGoogle Scholar
  4. 4.
    Sinden RE et al (1985) The development of Plasmodium ookinetes in vitro: an ultrastructural study including a description of meiotic division. Parasitology 91:227–244PubMedCrossRefGoogle Scholar
  5. 5.
    Dinglasan RR et al (2007) Plasmodium falciparum ookinetes require mosquito midgut chondroitin sulfate proteoglycans for cell invasion. Proc Natl Acad Sci USA 104:15882–15887PubMedCrossRefGoogle Scholar
  6. 6.
    Ghosh AK et al (2010) An improved method for the in vitro differentiation of Plasmodium falciparum gametocytes into ookinetes. Malar J 9:194–200PubMedCrossRefGoogle Scholar
  7. 7.
    Bounkeua V et al (2010) In vitro generation of Plasmodium falciparum ookinetes. Am J Trop Med Hyg 83:1187–1194PubMedCrossRefGoogle Scholar
  8. 8.
    McClean CM et al (2010) Optimized in vitro production of Plasmodium vivax ookinetes. Am J Trop Med Hyg 83:1183–1186PubMedCrossRefGoogle Scholar
  9. 9.
    Trager W, Jensen JB (1976) Human malaria parasites in continuous culture. Science 193:674–675CrossRefGoogle Scholar
  10. 10.
    Carter R, Miller LH (1979) Evidence for environmental modulation of gametocytogenesis in Plasmodium falciparum continuous culture. Bull World Health Organ 57:37–52PubMedGoogle Scholar
  11. 11.
    Billker O et al (1997) The roles of temperature, pH and mosquito factors as triggers of male and female gametogenesis of Plasmodium berghei in vitro. Parasitology 115:1–7PubMedCrossRefGoogle Scholar
  12. 12.
    Billker O et al (1998) Identification of xanthurenic acid as the putative inducer of malaria development in the mosquito. Nature 392:289–292PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Molecular Microbiology and ImmunologyJohns Hopkins University School of Public HealthBaltimoreUSA

Personalised recommendations