Biomechanical Characterization of Human Pluripotent Stem Cell-Derived Cardiomyocytes by Use of Atomic Force Microscopy.
Methods in Molecular Biology. 2018-10-30; : 343-353
DOI: 10.1007/978-1-4939-8894-5_20
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Pribyl J(1), Pešl M(2)(3)(4), Caluori G(1)(3), Acimovic I(2), Jelinkova S(2), Dvorak P(2)(3), Skladal P(1), Rotrekl V(5)(6).
Author information:
(1)CEITEC MU, Masaryk University, Brno, Czech Republic.
(2)Faculty of Medicine, Department of Biology, Masaryk University, Brno, Czech Republic.
(3)ICRC, St. Anne’s University Hospital, Brno, Czech Republic.
(4)Faculty of Medicine, First Department of Internal Medicine-Cardioangiology, Masaryk University, Brno, Czech Republic.
(5)Faculty of Medicine, Department of Biology, Masaryk University, Brno, Czech
Republic. .
(6)ICRC, St. Anne’s University Hospital, Brno, Czech Republic. .
Atomic force microscopy (AFM) is not only a high-resolution imaging technique but
also a sensitive tool able to study biomechanical properties of bio-samples
(biomolecules, cells) in native conditions-i.e., in buffered solutions (culturing
media) and stable temperature (mostly 37 °C). Micromechanical transducers
(cantilevers) are often used to map surface stiffness distribution, adhesion
forces, and viscoelastic parameters of living cells; however, they can also be
used to monitor time course of cardiomyocytes contraction dynamics (e.g. beating
rate, relaxation time), together with other biomechanical properties. Here we
describe the construction of an AFM-based biosensor setup designed to study the
biomechanical properties of cardiomyocyte clusters, through the use of standard
uncoated silicon nitride cantilevers. Force-time curves (mechanocardiograms, MCG)
are recorded continuously in real time and in the presence of
cardiomyocyte-contraction affecting drugs (e.g., isoproterenol, metoprolol) in
the medium, under physiological conditions. The average value of contraction
force and the beat rate, as basic biomechanical parameters, represent
pharmacological indicators of different phenotype features. Robustness, low
computational requirements, and optimal spatial sensitivity (detection limit
200 pN, respectively 20 nm displacement) are the main advantages of the presented
method.