TY - JOUR
T1 - Use of the MTT assay in adult ventricular cardiomyocytes to assess viability
T2 - Effects of adenosine and potassium on cellular survival
AU - Gomez, Luis A.
AU - Alekseev, Alexey E.
AU - Aleksandrova, Luba A.
AU - Brady, Peter A.
AU - Terzic, Andre
N1 - Funding Information:
This work was supported by the Pharmaceutical Research and Manufacturers of America Foundation, the American Heart Association, the Miami Heart Research Institute, and the Harrington Professorship Fund (to A.T.). The support that L.A.G. received from COLCIENCIAS and INS is acknowledged. P.A.B. is a recipient of a General Mills Clinician-Investigator Fellowship.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1997/4
Y1 - 1997/4
N2 - This study used the colorimetric MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)] assay to assess cell viability in isolated quiescent adult guinea-pig ventricular myocytes exposed to different insults or cardioprotective conditions, including adenosine and hyperkalemic-cardioplegia. Optical density (OD), reflecting intracellular reduction of MTT into formazan pigment formation, was a function of the number of viable cells (coefficient of linear correlation ~0.99), with MTT reduction preferentially carried out by rod-shaped cardiomyocytes (absorbance at 1.009 ± 0.013 and 0.006 ± 0.001 OD units for populations containing 50 and 0% of rod-shaped cells). Following prolonged mechanical (pressure of 1 lb/min for 40 min), chemical (10% DMSO or ethanol) or hypoxic injury (N2-saturated solution), the MTT reductase activity reflected reduction in the number of viable cells by 87%, > 50%, and 77%, respectively. In cardiomyocytes exposed to a 40 min hypoxia (with CO2), the MTT reductase activity was 0.056 ± 0.009 in the absence, and 0.074 ± 0.008 OD units in the presence of adenosine (1 mM), i.e. adenosine reduced the number of non-viable cells. Also, the MTT assay revealed that the effect of potassium-containing solutions (16 and 32 mMK+) on cellular viability may depend on the extent of insult imposed on cardiomyocytes; i.e. a ~24% and 49% increase under mild hypoxia (0.03% CO2), or an 18% decrease in cell viability under severe hypoxia (N2) in pre-injured cells. Thus, the MTT assay used to assess viability of isolated adult cardiomyocytes revealed a direct cytoprotective effect of adenosine and hyperkalemic-cardioplegia by promoting cell survival under certain conditions in vitro.
AB - This study used the colorimetric MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)] assay to assess cell viability in isolated quiescent adult guinea-pig ventricular myocytes exposed to different insults or cardioprotective conditions, including adenosine and hyperkalemic-cardioplegia. Optical density (OD), reflecting intracellular reduction of MTT into formazan pigment formation, was a function of the number of viable cells (coefficient of linear correlation ~0.99), with MTT reduction preferentially carried out by rod-shaped cardiomyocytes (absorbance at 1.009 ± 0.013 and 0.006 ± 0.001 OD units for populations containing 50 and 0% of rod-shaped cells). Following prolonged mechanical (pressure of 1 lb/min for 40 min), chemical (10% DMSO or ethanol) or hypoxic injury (N2-saturated solution), the MTT reductase activity reflected reduction in the number of viable cells by 87%, > 50%, and 77%, respectively. In cardiomyocytes exposed to a 40 min hypoxia (with CO2), the MTT reductase activity was 0.056 ± 0.009 in the absence, and 0.074 ± 0.008 OD units in the presence of adenosine (1 mM), i.e. adenosine reduced the number of non-viable cells. Also, the MTT assay revealed that the effect of potassium-containing solutions (16 and 32 mMK+) on cellular viability may depend on the extent of insult imposed on cardiomyocytes; i.e. a ~24% and 49% increase under mild hypoxia (0.03% CO2), or an 18% decrease in cell viability under severe hypoxia (N2) in pre-injured cells. Thus, the MTT assay used to assess viability of isolated adult cardiomyocytes revealed a direct cytoprotective effect of adenosine and hyperkalemic-cardioplegia by promoting cell survival under certain conditions in vitro.
KW - Adenosine
KW - Cardiomyocyte
KW - Cardioprotection
KW - DNP
KW - Hypoxia
KW - K
KW - MTT assay
KW - Viability
UR - http://www.scopus.com/inward/record.url?scp=0031127381&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031127381&partnerID=8YFLogxK
U2 - 10.1006/jmcc.1996.0363
DO - 10.1006/jmcc.1996.0363
M3 - Article
C2 - 9160877
AN - SCOPUS:0031127381
SN - 0022-2828
VL - 29
SP - 1255
EP - 1266
JO - Journal of Molecular and Cellular Cardiology
JF - Journal of Molecular and Cellular Cardiology
IS - 4
ER -