Folia Parasitologica 66:016 (2019) | DOI: 10.14411/fp.2019.016

Measuring mitochondrial respiration in adherent cells infected with Trypanosoma cruzi Chagas, 1909 using Seahorse extracellular flux analyser

Laura Maria González-Ortiz*, Juana Patricia Sánchez-Villamil*, Mike A. Celis-Rodríguez, Giovanni Lineros, Sandra Sanabria-Barrera, Norma C. Serrano, Melvin Y. Rincon, Paula K. Bautista-Nino
Traslational Biomedical Research Group, Centro de Investigaciones, Fundación Cardiovascular de Colombia, Santander, Colombia.
These authors contributed equally to this work

Infection with Trypanosoma cruzi Chagas, 1909 is reported to increase the production of reactive oxygen species in patients with Chagas disease. Mitochondria dysfunction, host inflammatory response and inadequate antioxidant response are described as the main factors leading to oxidative stress during acute and chronic stages of the disease. The Seahorse XFe24 extracellular flux platform allows energy metabolism determination through mitochondrial respiration and glycolysis measurements. XFe24 platform can be used in in vitro models of T. cruzi-infected cells, which allow the assessment and even modulation of endogenous conditions of infected cells, generating readouts of real-time cellular bioenergetics changes. In this protocol, we standardised the use of XFe24 technology in T. cruzi infected AC16 cardiomyocytes and SGHPL-5 trophoblasts. In addition, we provide a list of optimised assay specifications, advantages and critical steps to be considered during the process. Cardiomyocytes and trophoblasts are attractive target cells to evaluate the metabolic environment in acute, chronic and congenital Chagas transmission scenarios.

Keywords: Chagas disease, trophoblastic cells, cardiomyocytes, cellular respiration, mitochondrial bioenergetics.

Received: September 19, 2019; Accepted: May 6, 2019; Published online: October 10, 2019  Show citation

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González-Ortiz, L.M., Sánchez-Villamil, J.P., Celis-Rodríguez, M.A., Lineros, G., Sanabria-Barrera, S., Serrano, N.C., Rincon, M.Y., & Bautista-Nino, P.K. (2019). Measuring mitochondrial respiration in adherent cells infected with Trypanosoma cruzi Chagas, 1909 using Seahorse extracellular flux analyser. Folia Parasitologica, 66, Article 2019.016. https://doi.org/10.14411/fp.2019.016
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