Abstract
A method is described for the isolation and purification of trypomastigotes and amastigotes ofTrypanosoma cruzi from cell cultures. L-A9, a transformed fibroblast cell line, and J774G8, a macrophage-like cell line of tumor origin, were used. Both cell lines were infected with bloodstream trypomastigotes ofT. cruzi, which one within host cells transform into dividing amastigotes. After 6–8 days infection the host cells ruptured, spontaneously liberating parasites into the culture medium. L-A9 cells liberated mainly trypomastigotes while J774G8 cells liberated amastigotes. The parasites were collected and purified by centrifugation in a gradient of metrizamide. The purity of the preparation as well as the morphology of the parasites and the host cells were analysed by electron microscopy.
Similar content being viewed by others
References
Carvalho RMG, Meirelles MNL, De Souza W (1981) Isolation of the intracellular stage ofTrypanosoma cruzi and its interaction with mouse macrophages in vitro. Infect Immun 33:546–554
Damigni G, Kiyotaki W, Soeller W, Sasada M, Peisach J, Bloom BR (1980) Macrophage variants in oxygen metabolism. J Exp Med 152:808–818
Gutteridge WE, Cover B, Gaborak M (1978) Isolation of blood and intracellular forms ofTrypanosoma cruzi from rats and other rodents and preliminary studies on their metabolism. Parasitology 76:159–176
Leon W, Villalta F, Queiroz T, Szarfman A (1979) Antibody-induced capping of the intracellular stage ofTrypanosoma cruzi. Infect Immun 26:1218–1220
Loures MA, Pimenta PFP, De Souza W (1980) Isolation of bloodstream trypomastigotes ofTrypanosoma cruzi by a gradient of metrizamide. J Parasitol 66:1058–1059
Ralph P, Prichard J, Cohn M (1975) Reticulum cell sarcoma: An effector cell in antibody-dependent cell-mediated immunity. J Immunol 114:898–905
Sanderson CJ, Thomas JA, Twomey CE (1980) The growth ofTrypanosoma cruzi in human diploid cells for the production of trypomastigotes. Parasitology 80:153–162
Schmatz DM, Murray PK (1981)Trypanosoma cruzi: Selective isolation of pure trypomastigotes from cultured muscle cells. J Parasitol 67:517–521
Villalta FV, Leon W (1979) Effect of purification by DEAE-cellulose column on infectivity ofTrypanosoma cruzi blood forms. J Parasitol 65:188–189
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
de Carvalho, T.U., de Souza, W. Separation of amastigotes and trypomastigotes ofTrypanosoma cruzi from cultured cells. Z. Parasitenkd. 69, 571–575 (1983). https://doi.org/10.1007/BF00926668
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00926668