TY - JOUR
T1 - Measuring mitochondrial respiration in adherent cells infected with Trypanosoma cruzi Chagas, 1909 using Seahorse extracellular flux analyser
AU - Gonzalez-Ortiz, Laura Maria
AU - Sanchez-Villamil, Juana Patricia
AU - Celis-Rodriguez, Mike A.
AU - Lineros, Giovanni
AU - Sanabria-Barrera, Sandra
AU - Serrano, Norma C.
AU - Rincon, Melvin Y.
AU - Bautista-Nino, Paula K.
PY - 2019/10/10
Y1 - 2019/10/10
N2 - 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.
AB - 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.
KW - Chagas disease
KW - cardiomyocytes
KW - cellular respiration
KW - mitochondrial bioenergetics.
KW - trophoblastic cells
UR - http://www.scopus.com/inward/record.url?scp=85073625748&partnerID=8YFLogxK
U2 - 10.14411/fp.2019.016
DO - 10.14411/fp.2019.016
M3 - Artículo Científico
C2 - 31631068
AN - SCOPUS:85073625748
SN - 1803-6465
VL - 66
JO - Folia parasitologica
JF - Folia parasitologica
ER -
