Laboratory techniques to obtain different forms of Trypanosoma cruzi: applications to wild-type and genetically modified parasites

Abstract

Nowadays, there are no simple techniques for mimicking in vitro the life cycle of the kinetoplasmtid Trypanosoma cruzi Chagas, 1909, causative agent of Chagas disease, especially for parasite strains maintained as epimastigotes for many years. In the present study, we propose a method for obtaining metacyclic trypomastigotes, which were capable of infecting mammalian cells by simply lowering pH media. The collected amastigotes and trypomastigotes were differentiated into epimastigotes closing T. cruzi life cycle in vitro. Metacyclogenesis rates and infectivity were enhanced in cycled parasites. Finally, using this method, we were able to infect cells with transgenic parasites obtaining trypomastigotes and amastigotes using a neomycin-resistant cell line.