Hostname: page-component-848d4c4894-4hhp2 Total loading time: 0 Render date: 2024-05-17T23:42:39.041Z Has data issue: false hasContentIssue false
  • English
  • Français

The growth of Trypanosoma cruzi in human diploid cells for the production of trypomastigotes

Published online by Cambridge University Press:  06 April 2009

C. J. Sanderson ,
Jennifer A. Thomas  and
Cheryl E. Twomey
C. J. Sanderson
Affiliation:
Transplantation Biology, Clinical Research Centre, Watford Road, Harrow, Middlesex, HA1 3UJ
Jennifer A. Thomas
Affiliation:
Transplantation Biology, Clinical Research Centre, Watford Road, Harrow, Middlesex, HA1 3UJ
Cheryl E. Twomey
Affiliation:
Transplantation Biology, Clinical Research Centre, Watford Road, Harrow, Middlesex, HA1 3UJ
Get access Rights & Permissions [Opens in a new window]

Summary

The use of human diploid cell lines of finite life for the in vitro production of Trypanosoma cruzi is described. Both MRC5 and WI38 cells release trypomastigotes with less than 5% amastigotes. This could form the basis for biochemical and immunological studies, which were previously limited by the problems of separating parasites from blood. By selecting the in vitro passage number of the parasite it is possible to select for either the broad or the slender forms of trypomastigotes, allowing comparative studies of these forms within a single strain of the parasite. It is also possible to isolate amastigotes by disrupting the cells before trypomastigotes appear, and separating them from cell debris with Metrizamide. It is shown that by incorporating [3H]uridine in the cell-culture medium, labelled trypomastigotes are obtained. The release of this label (putative RNA) provides a relatively simple isotopic assay for parasite death. Examples of this assay for testing drug toxicity and in immunological lysis are presented.

Type
Research Article
Information
Parasitology , Volume 80 , Issue 1 , February 1980 , pp. 153 - 162
Copyright
Copyright © Cambridge University Press 1980

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

Brener, Z. (1976). Significance of morphologic variation of bloodstream forms. In New Approaches in American Trypanosomiasis Research. Scientific Publication no. 318, Pan American Health Organisation, Washington, pp. 127–31.Google Scholar
Budzko, D. B. & Kierszenbaum, F. (1974). Isolation of Trypanosoma cruzi from blood. Journal of Parasitology 60, 1037–8.CrossRefGoogle ScholarPubMed
Dvorak, J. A. (1976). New in vitro approach to quantitation of Trypanosoma cruzi vertebrate cell interactions. In New Approaches in American Trypanosomiasis Research. Scientific publication no. 318, Pan American Health Organisation, Washington, pp. 109–120.Google Scholar
Dvorak, J. A. & Hyde, T. P. (1973). Trypanosoma cruzi: interaction with vertebrate cells in vitro. I. Individual interactions at the cellular and subcellular levels. Experimental Parasitology 34, 268–83.CrossRefGoogle ScholarPubMed
Gutteridge, W. E., Cover, B. & Gaborak, M. (1978). Isolation of blood and intracellular forms of Trypanosoma cruzi from rats and other rodents and preliminary studies of their metabolism. Parasitology 76, 159–76.CrossRefGoogle ScholarPubMed
Gutteridge, W. E., Knowler, J. & Coombs, J. D. (1969). Growth of Trypanosoma cruzi in human heart tissue cells and effects of aminonucleoside of puromycin, trypacidin and aminopterin. Journal of Protozoology 16, 521–5.CrossRefGoogle ScholarPubMed
Hanson, W. L., Chapman, W. L. & Waits, V. B. (1976). Immunization of mice with irradiated Trypanosoma cruzi grown in cell culture: relation of numbers of parasites, immunizing injections and route of immunization to resistance. International Journal for Parasitology 6, 341–7.CrossRefGoogle Scholar
Jacobs, J. P., Jones, C. M. & Bailie, J. P. (1970). Characteristics of a human diploid cell designated MRC-5. Nature, London 227, 168–70.CrossRefGoogle ScholarPubMed
Kierszenbaum, F., Ivanyi, J. & Budzko, D. B. (1976). Mechanisms of natural resistance to trypanosomal infection. Role of complement in avian resistance to Trypanosoma cruzi infection. Immunology 30, 1–6.Google ScholarPubMed
Kofoid, C. A., Wood, F. C. & McNeil, E. (1935). The cycle of Trypanosoma cruzi in tissue culture of embryonic heart muscle. University of California Publications in Zoology 41, 23–4.Google Scholar
Lopez, A. F., Bunn Moreno, M. M. & Sanderson, C. J. (1979). The lysis of Trypanosoma cruzi epimastigotes by eosinophils and neutrophils. International Journal for Parasitology 8, 485–9.CrossRefGoogle Scholar
Maria, T. A., Tafuri, W. & Brener, Z. (1972). The fine structure of different bloodstream forms of Trypanosoma cruzi. Annals of Tropical Medicine and Parasitology 66, 423–31.CrossRefGoogle ScholarPubMed
Meyer, H. & Xavier de Oliveira, M. (1948). Cultivation of Trypanosoma cruzi in tissue cultures: a four year study. Parasitology 39, 91–4.CrossRefGoogle Scholar
Nogueira, N., Bianco, C. & Cohn, Z. (1975). Studies on the selective lysis and purification of Trypanosoma cruzi. Journal of Experimental Medicine 142, 224–9.CrossRefGoogle ScholarPubMed
Rodriguez, E. & Marinkelle, C. J. (1970). Trypanosoma cruzi: development in tissue culture. Experimental Parasitology 27, 78–87.CrossRefGoogle ScholarPubMed
Sanderson, C. J., Bunn Moreno, M. M. & Lopez, A. F. (1978). Antibody-dependent cell mediated cytotoxicity of Trypanosoma cruzi: the release of tritium labelled RNA, DNA and protein. Parasitology 76, 299–307.CrossRefGoogle ScholarPubMed
Sanderson, C. J. & de Souza, W. (1979). A morphological study of the interaction between Trypanosoma cruzi and rat eosinophils, neutrophils and macrophages in vitro. Journal of Cell Science 37, 275–86.CrossRefGoogle ScholarPubMed
Sanderson, C. J. & Thomas, J. A. (1978). A comparison of the cytotoxic activity of eosinophils and other cells by 51chromium release and time lapse microcinematography. Immunology 34, 771–80.Google Scholar