Laboratory techniques to obtain different forms of Trypanosoma cruzi: applications to wild-type and genetically modified parasites.
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.
Laboratorio de Biología Molecular de Trypanosoma cruzi, Instituto de Investigaciones Médicas 'Alfredo Lanari', Universidad de Buenos Aires and CONICET, Buenos Aires, Argentina.
María de los Milagros
Mde L
de Cámara
Laboratorio de Biología Molecular de Trypanosoma cruzi, Instituto de Investigaciones Médicas 'Alfredo Lanari', Universidad de Buenos Aires and CONICET, Buenos Aires, Argentina.
León A
LA
Bouvier
Laboratorio de Biología Molecular de Trypanosoma cruzi, Instituto de Investigaciones Médicas 'Alfredo Lanari', Universidad de Buenos Aires and CONICET, Buenos Aires, Argentina.
Mariana R
MR
Miranda
Laboratorio de Biología Molecular de Trypanosoma cruzi, Instituto de Investigaciones Médicas 'Alfredo Lanari', Universidad de Buenos Aires and CONICET, Buenos Aires, Argentina.
Chantal
C
Reigada
Laboratorio de Biología Molecular de Trypanosoma cruzi, Instituto de Investigaciones Médicas 'Alfredo Lanari', Universidad de Buenos Aires and CONICET, Buenos Aires, Argentina.
Claudio A
CA
Pereira
D000818:Animals; D002522:Chlorocebus aethiops; D008018:Life Cycle Stages; D030781:Organisms, Genetically Modified; D014349:Trypanosoma cruzi / Q000235:genetics* / Q000502:physiology*; D014709:Vero Cells