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Cultivation of Trypanosoma cruzi in irradiated muscle cells: improved synchronization and enhanced trypomastigote production

Published online by Cambridge University Press:  06 April 2009

D. M. Schmatz  and
P. K. Murray
D. M. Schmatz
Affiliation:
Department of Immunoparasitology, Merck Institute for Therapeutic Research, Rahway, New Jersey 07065
P. K. Murray
Affiliation:
Department of Immunoparasitology, Merck Institute for Therapeutic Research, Rahway, New Jersey 07065
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Summary

Since the in vitro production of trypomastigote stages of Trypanosoma cruzi in cell culture is frequently limited by (1) host cell overgrowth and (2) by the unequal redistribution of parasites after cell division resulting in asynchronous release of trypomastigotes, a culture system was devised in which host cell mitosis was inhibited by irradiation prior to parasite infection. L-6 rat myoblast cells when exposed to 3000 rad. of gamma radiation lost their ability to divide but remained susceptible to infection with, and capable of supporting the intracellular growth of, T. cruzi. Using this approach it proved possible to have virtually 100% of cells infected and achieve much better synchronization of trypomastigote release than with conventional culture systems. Additionally, the total number of parasites provided approached 1 × 109 trypomastigotes/ 150 cm2 flask, a significant increase over other culture systems. Preliminary studies with Plasmodium fallax and Eimeria tenella indicate that irradiated host cells may be utilized to advantage for the cultivation of other intracellular protozoa.

Type
Research Article
Information
Parasitology , Volume 85 , Issue 1 , August 1982 , pp. 115 - 125
Copyright
Copyright © Cambridge University Press 1982

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References

Bertelli, M. S. M. & Brener, Z. (1980). Infection of tissue culture cells with bloodstream trypomastigotes of Trypanosoma cruzi. Journal of Parasitology 66, 992–7.CrossRefGoogle ScholarPubMed
Camargo, E. P. (1964). Growth and differentiation in Trypanosoma cruzi: I. Origin of metacyclic trypanosomes in liquid media. Revista do Institute de Medicina tropicale de Sao Paulo 6, 93–100.Google ScholarPubMed
Crane, M. St. J. & Dvorak, J. A. (1979). Trypanosoma cruzi: Interaction with vertebrate cells. DNA synthesis and growth of intracellular amastigotes and their relationship to host cell DNA synthesis and growth. Journal of Protozoology 26, 599–604.CrossRefGoogle ScholarPubMed
Dvorak, J. A. (1976). New in vitro approach to quantitation of Trypanosoma crwzi-vertebrate cell interactions. In New Approaches in American Trypanosomiasis Research. Scientific publication no. 318, Chagas Disease. Scientific publication no. 347. Pan American Health Organization, Washington, pp. 1–10.Google Scholar
Dvorak, J. A. & Howe, C. L. (1976). The attraction of Trypanosoma cruzi to vertebrate cells in vitro. Journal of Protozoology 23, 534–7.CrossRefGoogle ScholarPubMed
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
Dvorak, J. A. & Poore, C. M. (1974). Trypanosoma cruzi: Interaction with vertebrate cells in vitro. IV. Environmental temperature effects. Experimental Parasitology 36, 150–7.CrossRefGoogle ScholarPubMed
Gavrilov, W., Lesneret, S. & Cowez, S. (1940). Emploi de la methode des cultures de tissus dans l'etude protozoaires les trypanosomes. Rivista Di Parasitologia, 4, 147–54.Google Scholar
Jessop, N. W. & Hay, R. J. (1979). Preparation, preservation, recovery and use of irradiated feeder layers in cell culture research. Tissue Culture Association Manual 5, 1137–9.CrossRefGoogle Scholar
Kofoid, C. A., Wood, F. D. & 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
Luban, N. A. & Dvorak, J. A. (1974). Trypanosoma cruzi: Interaction with vertebrate cells in vitro. III. Selection for biological characteristics following intracellular passage. Experimental Parasitology 36, 143–9.CrossRefGoogle Scholar
Neva, F. A., Malone, M. F. & Myers, B. R. (1961). Factors influencing the intracellular growth of Trypanosoma cruzi in vitro. American Journal of Tropical Medicine and Hygiene 10, 140–54.CrossRefGoogle ScholarPubMed
Pan, S. C.-T. (1978). Trypanosoma cruzi: In vitro interactions between cultured amastigotes and human skin-muscle cells. Experimental Parasitology 45, 274–86.CrossRefGoogle ScholarPubMed
Puck, T. T. & Marcus, P. I. (1955). A rapid method for viable cell titration and clone production with HeLa cells in tissue culture: The use of X-irradiated cells to supply conditioning factors. Proceedings of the National Academy of Sciences U.S.A. 41, 432–7.CrossRefGoogle ScholarPubMed
Sanderson, C. J., Thomas, J. A. & Twomey, C. E. (1980). The growth of Trypanosoma cruzi in human diploid cells for the production of trypomastigotes. Parasitology 80, 153–62.CrossRefGoogle ScholarPubMed
Schmatz, D. M. & Murray, P. K. (1981). Selective isolation of pure trypomastigotes from cultured muscle cells. Journal of Parasitology 67, 517–21.CrossRefGoogle ScholarPubMed
Silva, L. H. P. & Nussenzweig, V. (1953). Sobre uma cepa de Trypanosoma cruzi altamente virulenta para o camundongo branco. Folia Clinical Biology 20, 191–207.Google Scholar
Trejos, A., Godoy, G. A., Greenblatt, C. & Cedillos, R. (1963). Effects of temperature on morphologic variation of Schizotrypanum cruzi in tissue culture. Experimental Parasitology 13, 211–18.CrossRefGoogle ScholarPubMed
Yaffe, D. (1971). Developmental changes preceding cell fusion during muscle differentiation in vitro. Experimental Cell Research 66, 33–48.CrossRefGoogle ScholarPubMed
Yaffe, D. & Feldmann, M. (1965). The formation of hybrid multinucleated muscle fibers from myoblasts of different genetic origin. Developmental Biology 11, 300–17.CrossRefGoogle ScholarPubMed