Separation of amastigotes and trypomastigotes of Trypanosoma cruzi from cultured cells.
A method is described for the isolation and purification of trypomastigotes and amastigotes of Trypanosoma 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 of T. cruzi, which once 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.
D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D005347:Fibroblasts; D008264:Macrophages; D008854:Microscopy, Electron; D014349:Trypanosoma cruzi / Q000166:cytology / Q000254:growth & development / Q000302:isolation & purification*