Folia Parasitologica, vol. 60 (2013), issue 5

Folia Parasitologica 60[5] 406-410 (2013) | DOI: 10.14411/fp.2013.042

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

María de los Milagros Cámara, León A. Bouvier, Mariana R. Miranda, Chantal Reigada, Claudio A. Pereira
Laboratorio de Biología Molecular de Trypanosoma cruzi, Instituto de Investigaciones Médicas 'Alfredo Lanari', Universidad de Buenos Aires and CONICET, Buenos Aires, Argentina

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.

Keywords: Chagas disease, metacyclogenesis, parasite differentiation, intracellular epimastigotes

Received: December 4, 2012; Accepted: May 6, 2013; Published: November 27, 2013


References

  1. Almeida-De-Faria M., Freymuller E., Colli W., Alves M.J. 1999: Trypanosoma cruzi: characterization of an intracellular epimastigote-like form. Exp. Parasitol. 92: 263-274 Go to original source... Go to PubMed...
  2. Camargo E.P. 1964: Growth and Differentiation in Trypanosoma cruzi. I. Origin of metacyclic trypanosomes in liquid media. Rev. Inst. Med. Trop. Sao Paulo. 6: 93-100 Go to PubMed...
  3. Castellani O., Ribeiro L.V., Fernandes J.F. 1967: Differentiation of Trypanosoma cruzi in culture. J. Protozool. 14: 447-451 Go to original source... Go to PubMed...
  4. Contreras V.T., Navarro M.C., De Lima A.R., Arteaga R., Duran F., Askue J., Franco Y. 2002: Production of amastigotes from metacyclic trypomastigotes of Trypanosoma cruzi. Mem. Inst. Oswaldo Cruz. 97: 1213-1220 Go to original source... Go to PubMed...
  5. Contreras V.T., Salles J.M., Thomas N., Morel C.M., Goldenberg S. 1985: In vitro differentiation of Trypanosoma cruzi under chemically defined conditions. Mol. Biochem. Parasitol. 16: 315-327 Go to original source... Go to PubMed...
  6. De Lima A.R., Navarro M.C., Arteaga R.Y., Contreras V.T. 2008: Cultivation of Trypanosoma cruzi epimastigotes in low glucose axenic media shifts its competence to differentiate at metacyclic trypomastigotes. Exp. Parasitol. 119: 336-342 Go to original source... Go to PubMed...
  7. Dusanic D.G. 1980: In vitro production of metacyclic trypomastigotes of Trypanosoma cruzi. J. Parasitol. 66: 1046-1049 Go to original source... Go to PubMed...
  8. Goldenberg S., Avila A.R. 2011: Aspects of Trypanosoma cruzi stage differentiation. Adv. Parasitol. 75: 285-305 Go to original source... Go to PubMed...
  9. Kimura E., Lay W.H., Fernandes J.F. 1978: Extracellular 'in vitro' evolution of metacyclic trypomastigotes isolated from Trypanosoma cruzi cultures. Rev. Inst. Med. Trop. Sao Paulo. 20: 133-138 Go to PubMed...
  10. Kolev N.G., Ramey-Butler K., Cross G.A., Ullu E., Tschudi C. 2012: Developmental progression to infectivity in Trypanosoma brucei triggered by an RNA-binding protein. Science 338: 1352-1353 Go to original source... Go to PubMed...
  11. Navarro M.C., De Lima A.R., Askue J., Contreras V.T. 2003: Morphological comparison of axenic amastigogenesis of trypomastigotes and metacyclic forms of Trypanosoma cruzi. Mem. Inst. Oswaldo Cruz. 98: 83-91 Go to original source... Go to PubMed...
  12. Pereira C.A., Alonso G.D., Ivaldi S., Silber A.M., Alves M.J., Torres H.N., Flawia M.M. 2003: Arginine kinase overexpression improves Trypanosoma cruzi survival capability. FEBS Lett. 554: 201-205 Go to original source... Go to PubMed...
  13. Rondinelli E., Silva R., Carvalho J.F., De Almeida Soares C.M., De Carvalho E.F., De Castro F.T. 1988: Trypanosoma cruzi: an in vitro cycle of cell differentiation in axenic culture. Exp. Parasitol. 66: 197-204 Go to original source... Go to PubMed...
  14. Steverding D. 2008: The history of African trypanosomiasis. Parasit. Vectors 1: 3 Go to original source... Go to PubMed...
  15. Tomlinson S., Vandekerckhove F., Frevert U., Nussenzweig V. 1995: The induction of Trypanosoma cruzi trypomastigote to amastigote transformation by low pH. Parasitology 110: 547-554 Go to original source... Go to PubMed...
  16. Ucros H., Granger B., Krassner S.M. 1983: Trypanosoma cruzi: effect of pH on in vitro formation of metacyclic trypomastigotes. Acta Trop. 40: 105-112 Go to PubMed...