Hostname: page-component-cd9895bd7-gvvz8 Total loading time: 0 Render date: 2024-12-26T00:27:41.006Z Has data issue: false hasContentIssue false

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

Published online by Cambridge University Press:  06 April 2009

R. E. Sinden
Affiliation:
Department of Pure and Applied Biology, Imperial College, London SW7 2EE
T. Ponnudurai
Affiliation:
Department of Medical Parasitology, Geert Grooteplein Zuid 24, University of Nijmegen, 6500 HB, Nijmegen, The Netherlands
M. A. Smits
Affiliation:
Department of Molecular Biology, University of Nijmegen, Toernooiveld, 6525ED, Nijmegen, The Netherlands
A. M. Simm
Affiliation:
Department of Pure and Applied Biology, Imperial College, London SW7 2EE
J. H. E. Th. Meuwissen
Affiliation:
Department of Medical Parasitology, Geert Grooteplein Zuid 24, University of Nijmegen, 6500 HB, Nijmegen, The Netherlands

Summary

A 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.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1984

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Brockleman, C. R. (1982). Conditions favouring gametocytogenesis in the continuous culture of Plasmodium falciparum. Journal of Protozoology 29, 454–8.Google Scholar
Hawking, F., Wilson, M. E. & Gammage, K. (1971). Evidence for the cyclic development and short lived maturity in gametocytes of Plasmodium falciparum. Transactions of the Royal Society of Tropical Medicine and Hygiene 65, 549–59.CrossRefGoogle ScholarPubMed
Hommel, M., Selkirk, M. & Miltgen, F. (1979). A method for the concentration of viable gametocytes of Plasmodium chabaudi. Transactions of the Royal Society of Tropical Medicine and Hygiene 73, 326–7.Google Scholar
Kass, L., Willerson, D. Jr., Riekmann, K. H., Carson, P. E. & Becker, R. F. (1971). Plasmodium falciparum gametocytes. Electron microscope observations on material obtained by a new method. American Journal of Tropical Medicine and Hygiene 20, 187–94.CrossRefGoogle ScholarPubMed
Kaushal, D. C., Carter, R., Miller, L. H. & Krishna, G. (1980). Gametocytogenesis by malaria parasites in continuous culture. Nature, London 286, 490–2.CrossRefGoogle ScholarPubMed
Knight, A. & Sinden, R. E. (1982). The purification of gametocytes of Plasmodium falciparum and P. yoelli nigeriensis by colloidal silica gradient centrifugation. Transactions of the Royal Society of Tropical Medicine and Hygiene 76, 503–9.CrossRefGoogle Scholar
Ponnudurai, T., Lensen, A. H. W., Leeuwenberg, A. D. E. M. & Meuwissen, J. H. E. Th. (1982). Cultivation of fertile Plasmodium falciparum gametocytes in semiautomated systems. I. Static cultures. Transactions of the Royal Society of Tropical Medicine and Hygiene 76, 812–18.CrossRefGoogle ScholarPubMed
Ponnudurai, T., Lensen, A. H. W. & Meuwissen, J. H. E. Th. (1983). An automated large-scale culture system of Plasmodium falciparum using tangential flow filtration for medium change. Parasitology 87, 439–45.CrossRefGoogle ScholarPubMed
Sinden, R. E. (1983). Sexual development of malarial parasites. Advances in Parasitology 22, 153216.CrossRefGoogle ScholarPubMed
Sinden, R. E. & Smalley, M. E. (1979). Gametocytogenesis of Plasmodium falciparum in vitro: the cell cycle. Parasitology 79, 277–96.Google Scholar
Smalley, M. E. (1977). Plasmodium falciparum gametocytes: the effect of chloroquine on their development. Transactions of the Royal Society of Tropical Medicine and Hygiene 75, 316–17.Google Scholar
Vermeulen, A., Ponnudurai, T., Lensen, A. H. W., Roeffen, W. F. G. & Meuwissen, J. H. E. Th. (1983). The purification of Plasmodium falciparum macrogametes and/or zygotes prepared from in vitro cultures. Transactions of the Royal Society of Tropical Medicine and Hygiene (in the Press).CrossRefGoogle ScholarPubMed
Williamson, J. & Cover, B. (1975). The rapid isolation from human blood of concentrated, white-cell-free preparations of Plasmodium falciparum. Transactions of the Royal Society of Tropical Medicine and Hygiene 69, 7887.CrossRefGoogle ScholarPubMed