Synchronization of Plasmodium falciparum gametocytes using an automated suspension culture system | Parasitology | Cambridge Core (original) (raw)

Summary

An automated suspension culture system for the cultivation of Plasmodium falciparum is described which retains a degree of flexibility which is absent in other automated culture apparatuses. Not only does this system of cultivation promote rapid multiplication of asexual parasites but also permits the development and maturation of gametocytes. Using a combination of gelatin flotation and _N_-acetyl glucosamine treatment synchronous development of gametocytes was achieved. The total time for gametocyte maturation in vitro under the conditions provided was 7 days. Stages II and V required 48 h for development whilst I, III and IV needed 24 h each. Mature microgametocytes were relatively long lived in comparison with macrogametocytes. Electron microscopic study of the synchronized stages confirmed the observations of Sinden (1982) but, in addition, we noted the presence of Garnham bodies, a cytostome in all stages and dense spherules in stages I–III similar to the fenestrated buttons in sporozoites and exoerythrocytic forms. The relationship between the number of osmiophilic bodies in the mature gametocytes and their ability to escape from the red cell is reaffirmed.

References

Butcher, G. A. (1981 a). A comparison of static thin layer and suspension cultures for the maintenance in vitro of Plasmodium falciparum. Annals of Tropical Medicine and Parasitology 75, 7–17.CrossRefGoogle ScholarPubMed

Butcher, G. A. (1981 b). A simple automated device for the maintenance of Plasmodium falciparum in suspension culture. Annals of Tropical Medicine and Parasitology 75, 111–13.CrossRefGoogle ScholarPubMed

Carter, R. & Beach, R. F. (1978). Gametogenesis in culture by gametocytes of Plasmodium falciparum. Nature, London 270, 240–1.CrossRefGoogle Scholar

Carter, R. & Miller, L. H. (1979). Evidence for environmental modulation of gametocytogenesis of Plasmodium falciparum in continuous culture. Bulletin of the World Health Organization 57 (Suppl.), 37–52.Google ScholarPubMed

Diggs, C. L., Aikawa, M. & Hayes, J. D. (1977). Ultrastructure and viability of cryopreserved Plasmodium falciparum. Bulletin of the World Health Organization 55, 299–304.Google ScholarPubMed

Garnham, P. C. C., Bird, R. G. & Baker, J. R. (1960). Electron microscope studies of motile stages of malarial parasites 1. The fine structure of the sporozoites of Plasmodium gallinaceum. Transactions of the Royal Society of Tropical Medicine and Hygiene 54, 274–8.CrossRefGoogle Scholar

Gupta, S. K., Schulman, S. & Vanderberg, J. P. (1985). Stage dependent toxicity of _N_-acetylglucosamine to Plasmodium falciparum. Journal of Protozoology 32, 91–5.CrossRefGoogle ScholarPubMed

Hawking, F., Wilson, M. E. & Gammage, K. (1971). Evidence for cyclic development and short-lived maturity in the gametocytes of Plasmodium falciparum. Transactions of the Royal Society of Tropical Medicine and Hygiene 65, 549–59.CrossRefGoogle ScholarPubMed

Ifediba, T. & Vanderberg, J. P. (1981). Complete in vitro maturation of Plasmodium falciparum gametocytes. Nature, London 294, 364–96.CrossRefGoogle ScholarPubMed

Jensen, J. B. (1978). Concentrations from continuous culture of erythrocytes infected with trophozoites and schizonts of Plasmodium falciparum. American Journal of Tropical Medicine and Hygiene 27, 1274–6.CrossRefGoogle ScholarPubMed

Jensen, J. B. (1979). Observations on gametogenesis in Plasmodium falciparum from continuous culture. Journal of Protozoology 26, 129–32.CrossRefGoogle ScholarPubMed

Meis, J. F. G. M., Rijntjes, P. J. M., Verhave, J. P., Ponnudurai, T., Hollingdale, M. R., Sinden, R. E., Smith, J. E., Jap, P. H. K., Meuwissen, J. H. E. Th. & Yap, S. H. (1986). Fine structure of the malaria parasite Plasmodium falciparum in human hepatocytes in vitro. Cell and Tissue Research 244, 345–50.CrossRefGoogle ScholarPubMed

Ponnudurai, T., Meuwissen, J. H. E. Th., Leeuwenberg, A. D. E. M., Verhave, J. P. & Lensen, A. H. W. (1982 a). The production of mature gametocytes of Plasmodium falciparum in continuous cultures of different isolates infective to mosquitoes. Transactions of the Royal Society of Tropical Medicine and Hygiene 76, 242–50.CrossRefGoogle ScholarPubMed

Ponnudurai, T., Lensen, A. H. W., Leeuwenberg, A. D. E. M. & Meuwissen, J. H. E. Th. (1982 b). Cultivation of fertile Plasmodium falciparum gametocytes in semi-automated systems. I. Static cultures. Transactions of the Royal Society of Tropical Medicine and Hygiene 76, 812–8.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. (1982). Gametocytogenesis of Plasmodium falciparum in vitro: an electron microscopic study. Parasitology 84, 1–11.CrossRefGoogle ScholarPubMed

Sinden, R. E. (1983). Sexual development of malaria parasites. In Advances in Parasitology vol. 22, (ed. Baker, J. R. and R., Muller), pp. 153–216. London: Academic Press.Google Scholar

Sinden, R. E., Canning, E. U., Bray, R. S. & Smalley, M. E. (1978). Gametocyte and gamete development in Plasmodium falciparum. Proceedings of the Royal Society of London, B 201, 375–99.Google ScholarPubMed

Sinden, R. E., Canning, E. U. & Spain, B. J. (1976). Gametogenesis and fertilization in Plasmodium yoelii nigeriensis: a transmission electron microscope study. Proceedings of the Royal Society of London, B 193, 55–76.Google ScholarPubMed

Sinden, R. E. & Smalley, M. E. (1979). Gametocytogenesis of Plasmodium falciparum in vitro: the cell cycle. Parasitology 79, 277–96.CrossRefGoogle ScholarPubMed

Smalley, M. E. & Sinden, R. E. (1977). Plasmodium falciparum gametocytes: their longevity and infectivity. Parasitology 74, 1–8.CrossRefGoogle ScholarPubMed

Vermuelen, A. N., van Deursen, J., Brakenhoff, R. H., Lensen, A. H. W., Ponnudurai, T. & Meuwissen, J. H. E. Th. (1986). Characterization of Plasmodium falciparum sexual stage antigens and biosynthesis in synchronized gametocyte cultures. Molecular and Biochemical Parasitology 19, (in the Press).CrossRefGoogle Scholar

Zolg, J. W., Macleod, A. J., Dickson, I. A. & Scaife, J. G. (1982). Plasmodium falciparum: modifications of the in vitro culture conditions improving parasite yields. Journal of Parasitology 68, 1072–80.CrossRefGoogle ScholarPubMed