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@article{1405767,
author = {Caluori, Guido and Přibyl, Jan and Cmiel, Vratislav and Pešl, Martin and Potocnak, Tomas and Provazník, Ivo and Skládal, Petr and Rotrekl, Vladimír},
article_location = {Hoboken},
article_number = {2},
doi = {http://dx.doi.org/10.1002/jmr.2760},
keywords = {calcium imaging; atomic force microscopy; human stem cell-derived cardiomyocytes; in vitro models; fluorescence microscopy; cardiac differentiation; caffeine; embryoid bodies; biosignals filtering},
language = {eng},
issn = {0952-3499},
journal = {Journal of Molecular Recognition},
title = {Simultaneous study of mechanobiology and calcium dynamics on hESC-derived cardiomyocytes clusters},
url = {http://dx.doi.org/10.1002/jmr.2760},
volume = {32},
year = {2019}
}
TY - JOUR
ID - 1405767
AU - Caluori, Guido - Přibyl, Jan - Cmiel, Vratislav - Pešl, Martin - Potocnak, Tomas - Provazník, Ivo - Skládal, Petr - Rotrekl, Vladimír
PY - 2019
TI - Simultaneous study of mechanobiology and calcium dynamics on hESC-derived cardiomyocytes clusters
JF - Journal of Molecular Recognition
VL - 32
IS - 2
SP - 1-9
EP - 1-9
PB - Wiley
SN - 09523499
KW - calcium imaging
KW - atomic force microscopy
KW - human stem cell-derived cardiomyocytes
KW - in vitro models
KW - fluorescence microscopy
KW - cardiac differentiation
KW - caffeine
KW - embryoid bodies
KW - biosignals filtering
UR - http://dx.doi.org/10.1002/jmr.2760
L2 - http://dx.doi.org/10.1002/jmr.2760
N2 - Calcium ions act like ubiquitous second messengers in a wide amount of cellular processes. In cardiac myocytes, Ca2+ handling regulates the mechanical contraction necessary to the heart pump function. The field of intracellular and intercellular Ca2+ handling, employing in vitro models of cardiomyocytes, has become a cornerstone to understand the role and adaptation of calcium signalling in healthy and diseased hearts. Comprehensive in vitro systems and cell-based biosensors are powerful tools to enrich and speed up cardiac phenotypic and drug response evaluation. We have implemented a combined setup to measure contractility and calcium waves in human embryonic stem cells-derived cardiomyocyte 3D clusters, obtained from embryoid body differentiation. A combination of atomic force microscopy to monitor cardiac contractility, and sensitive fast scientific complementary metal-oxide-semiconductor camera for epifluorescence video recording, provided correlated signals in real time. To speed up the integrated data processing, we tested several post-processing algorithms, to improve the automatic detection of relevant functional parameters. The validation of our proposed method was assessed by caffeine stimulation (10mM) and detection/characterization of the induced cardiac response. We successfully report the first simultaneous recording of cardiac contractility and calcium waves on the described cardiac 3D models. The drug stimulation confirmed the automatic detection capabilities of the used algorithms, measuring expected physiological response, such as elongation of contraction time and Ca2+ cytosolic persistence, increased calcium basal fluorescence, and transient peaks. These results contribute to the implementation of novel, integrated, high-information, and reliable experimental systems for cardiac models and drug evaluation.
ER -
CALUORI, Guido, Jan PŘIBYL, Vratislav CMIEL, Martin PEŠL, Tomas POTOCNAK, Ivo PROVAZNÍK, Petr SKLÁDAL and Vladimír ROTREKL. Simultaneous study of mechanobiology and calcium dynamics on hESC-derived cardiomyocytes clusters. \textit{Journal of Molecular Recognition}. Hoboken: Wiley, 2019, vol.~32, No~2, p.~1-9. ISSN~0952-3499. Available from: https://dx.doi.org/10.1002/jmr.2760.
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