Human engineered heart tissue as a versatile tool in basic research and preclinical toxicology.
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Human engineered heart tissue as a versatile tool in basic research and preclinical toxicology. / Schaaf, Sebastian; Shibamiya, Aya; Mewe, Marco; Eder, Alexandra; Stöhr, Andrea; Hirt, Marc; Rau, Thomas; Zimmermann, Wolfram-Hubertus; Conradi, Lenard; Eschenhagen, Thomas; Hansen, Arne.
in: PLOS ONE, Jahrgang 6, Nr. 10, 10, 2011, S. 26397.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Human engineered heart tissue as a versatile tool in basic research and preclinical toxicology.
AU - Schaaf, Sebastian
AU - Shibamiya, Aya
AU - Mewe, Marco
AU - Eder, Alexandra
AU - Stöhr, Andrea
AU - Hirt, Marc
AU - Rau, Thomas
AU - Zimmermann, Wolfram-Hubertus
AU - Conradi, Lenard
AU - Eschenhagen, Thomas
AU - Hansen, Arne
PY - 2011
Y1 - 2011
N2 - Human embryonic stem cell (hESC) progenies hold great promise as surrogates for human primary cells, particularly if the latter are not available as in the case of cardiomyocytes. However, high content experimental platforms are lacking that allow the function of hESC-derived cardiomyocytes to be studied under relatively physiological and standardized conditions. Here we describe a simple and robust protocol for the generation of fibrin-based human engineered heart tissue (hEHT) in a 24-well format using an unselected population of differentiated human embryonic stem cells containing 30-40% ?-actinin-positive cardiac myocytes. Human EHTs started to show coherent contractions 5-10 days after casting, reached regular (mean 0.5 Hz) and strong (mean 100 µN) contractions for up to 8 weeks. They displayed a dense network of longitudinally oriented, interconnected and cross-striated cardiomyocytes. Spontaneous hEHT contractions were analyzed by automated video-optical recording and showed chronotropic responses to calcium and the ?-adrenergic agonist isoprenaline. The proarrhythmic compounds E-4031, quinidine, procainamide, cisapride, and sertindole exerted robust, concentration-dependent and reversible decreases in relaxation velocity and irregular beating at concentrations that recapitulate findings in hERG channel assays. In conclusion this study establishes hEHT as a simple in vitro model for heart research.
AB - Human embryonic stem cell (hESC) progenies hold great promise as surrogates for human primary cells, particularly if the latter are not available as in the case of cardiomyocytes. However, high content experimental platforms are lacking that allow the function of hESC-derived cardiomyocytes to be studied under relatively physiological and standardized conditions. Here we describe a simple and robust protocol for the generation of fibrin-based human engineered heart tissue (hEHT) in a 24-well format using an unselected population of differentiated human embryonic stem cells containing 30-40% ?-actinin-positive cardiac myocytes. Human EHTs started to show coherent contractions 5-10 days after casting, reached regular (mean 0.5 Hz) and strong (mean 100 µN) contractions for up to 8 weeks. They displayed a dense network of longitudinally oriented, interconnected and cross-striated cardiomyocytes. Spontaneous hEHT contractions were analyzed by automated video-optical recording and showed chronotropic responses to calcium and the ?-adrenergic agonist isoprenaline. The proarrhythmic compounds E-4031, quinidine, procainamide, cisapride, and sertindole exerted robust, concentration-dependent and reversible decreases in relaxation velocity and irregular beating at concentrations that recapitulate findings in hERG channel assays. In conclusion this study establishes hEHT as a simple in vitro model for heart research.
KW - Humans
KW - Reproducibility of Results
KW - Fluorescent Antibody Technique
KW - Cell Differentiation
KW - HEK293 Cells
KW - Embryonic Stem Cells/cytology
KW - RNA, Messenger/genetics/metabolism
KW - Arrhythmias, Cardiac/chemically induced
KW - Drug Evaluation, Preclinical/methods
KW - Electrophysiological Processes/drug effects
KW - Heart/drug effects/physiology
KW - Muscle Contraction/drug effects
KW - Myocardium/cytology/metabolism
KW - Myocytes, Cardiac/cytology/drug effects/metabolism
KW - Tissue Engineering
KW - Toxicity Tests/methods
KW - Humans
KW - Reproducibility of Results
KW - Fluorescent Antibody Technique
KW - Cell Differentiation
KW - HEK293 Cells
KW - Embryonic Stem Cells/cytology
KW - RNA, Messenger/genetics/metabolism
KW - Arrhythmias, Cardiac/chemically induced
KW - Drug Evaluation, Preclinical/methods
KW - Electrophysiological Processes/drug effects
KW - Heart/drug effects/physiology
KW - Muscle Contraction/drug effects
KW - Myocardium/cytology/metabolism
KW - Myocytes, Cardiac/cytology/drug effects/metabolism
KW - Tissue Engineering
KW - Toxicity Tests/methods
U2 - 10.1371/journal.pone.0026397
DO - 10.1371/journal.pone.0026397
M3 - SCORING: Journal article
VL - 6
SP - 26397
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 10
M1 - 10
ER -
