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Growth of Trypanosoma cruzi in vitro: development and application of a continuous-flow culture system

Published online by Cambridge University Press:  06 April 2009

G. T. Williams
Affiliation:
Department of Immunology, St George's Hospital Medical School, London SW17 0RE
L. Hudson
Affiliation:
Department of Immunology, St George's Hospital Medical School, London SW17 0RE

Summary

The design and operation of a modular, bacteriological continuous-flow culture system have been adapted for the growth of Trypanosoma cruzi epimastigotes in simple monophasic media. The system was designed to achieve a minimum of 200 days of continuous culture and provision was made for the continuous supply of medium and collection of parasites under sterile conditions. The system provides large quantities of epimastigotes with homogeneous morphology and uniform viability. The system also lends itself tothe analysis of the factors which affect parasite growth. We have examined the effects of changes in environmental parameters on epimastigote growth rate. Optimal growth was observed at 27 °C. The rate ofstirring of the culture had a small but definable effect on the growth rate, which was greatest at 80 r.p.m. Growth was only slightly affected by the level of dissolved oxygen between 10 and 50% saturation, but was inhibited at higher concentrations. Growth was slower at extreme values of pH but showed a broad optimum around pH 7·4.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1982

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References

Acton, R. T., Barstad, P. A., & Zwerner, R. K., (1979). Propagation and scaling-up of suspension cultures. Methods in Enzymology 58, 211–21.CrossRefGoogle ScholarPubMed
Avila, J. L., Bretana, A., Casanova, M. A., Avila, A., & Rodriguez, F., (1979). Trypanosoma cruzi: defined medium for continuous cultivation of virulent parasites. Experimental Parasitology 48, 2735.CrossRefGoogle ScholarPubMed
Avila, J. L., Casanova, M. A., Avila, A., & Bhetana, A., (1979). Acid and neutral hydrolases in Trypanosomn cruzi. Characterisation and assay. Journal of Protozoology 26, 304–11.CrossRefGoogle ScholarPubMed
Boné, G. J., & Parent, G., (1963). Stearic acid, an essential growth factor for Trypanosoma cruzi. Journal of General Microbiology 31, 261–6.CrossRefGoogle ScholarPubMed
Camargo, E. P., (1964). Growth and differentiation in Trypanosoma cruzi. I. Origin of metacyclic trypanosomes in liquid media. Revista do Instituto da Medicina Tropical de São Paulo 6, 93100.Google ScholarPubMed
Chang, S. L., (1947). Studies on haemoflagellates. I. A semi-solid medium and a fluid medium with a solid base for growing various species of Leishmania and Trypanosoma cruzi. Journal of Infectious Diseases 80, 164–71.CrossRefGoogle Scholar
Cross, G. A. M., Klein, R. A., & Baker, J. R., (1975). Trypanosoma cruzi: growth, amino acid utilisation and drug action in a defined medium. Annals of Tropical Medicine and Parasitology 69, 513–14.CrossRefGoogle Scholar
Fernandes, J. F., & Castellani, O., (1966). Growth characteristics and chemical composition of Trypanosoma cruzi. Experimental Parasitology 18, 195202.CrossRefGoogle Scholar
Gutteridge, W. E., (1981). Biochemistry of Trypanosoma cruzi. Transactions of the Royal Society of Tropical Medicine and Hygiene 75, 484–92.CrossRefGoogle Scholar
Gutteridge, W. E., & Rogerson, G. W., (1979). Biochemical aspects of the biology of Trypanosoma cruzi. In Biology of the Kinetoplastids (ed.Lumsden, W. R. H. and Evans, D. A.) pp. 619652. Vol. 2, London: Academic Press.Google Scholar
Hirtenstein, M., Clark, J., Lindgren, G., & Vretblad, P., (1979). Microcarriers of animal cell culture: a brief review of theory and practice. In Developments in Biological Standards. Proceedings of the 3rd General Meeting of the U.K. Society of Animal Cell Technology,3rd General MeetingOxford.October 1979, pp. 109116. Basel: S. Karger.Google Scholar
Itow, S., & Plessmann Camargo, E., (1977). Proteolytic activities in cell extracts of Trypanosoma cruzi. Journal of Protozoology 24, 591–5.CrossRefGoogle ScholarPubMed
McLimans, W. F., (1979). Mass culture of mammalian cells. Methods in Enzymology 58, 194211.CrossRefGoogle ScholarPubMed
Pereira da Silva, L. H., & Nussenzweig, V., (1953). Sobre uma cepa de Trypanosoma cruzi altamente virulenta para o camundongo branco. Folia Clinica Biologica São Paulo 20, 191208.Google Scholar
Phelps, A., (1936). Growth of Protozoa in pure culture. Journal of Experimental Zoology 72, 479–96.CrossRefGoogle Scholar
Pirt, S. J., (1975). Principles of Microbe and Cell Cultivation. London: Blackwell Scientific Publications.Google Scholar
Pirt, S. J., & Callow, D. S., (1959). Continuous-flow culture of the filamentous mould Penicillium chrysogenum and the control of its morphology. Nature, London 184, 307–10.CrossRefGoogle ScholarPubMed
Senekjie, H. A., (1943). Biochemical reactions, cultural characteristics and growth requirements of Trypanosoma cruzi. American Journal of Tropical Medicine 23, 523–31.Google Scholar
Snary, D., & Hudson, L., (1979). Trypanosoma cruzi cell surface proteins: identification of one major glycoprotein. FEBS Letters 100, 166–70.CrossRefGoogle ScholarPubMed
Tovey, M., & Bronty-Boyé, D., (1976). Characteristics of the chemostat culture of murine leukaemia L1210 cells. Experimental Cell Research 101, 346–54.CrossRefGoogle Scholar
Violaga, F. C., Roberts, E., & Penn, I., (1966). The effect of hyperbaric oxygen on aerobic bacteria. I. in vitro studies. Canadian Journal of Microbiology 12, 521–9.Google Scholar
Warren, L. G., (1960). Metabolism of Schizotrypanum cruzi. Journal of Parasitology 46, 529–39.CrossRefGoogle ScholarPubMed