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Study of mechanobiological properties of human functional cardiomyocytes using Atomic Force Microscopy

Název česky Výzkum mechanobiologických vlastností lidských funkčních kardiomyocytů s využitím Mikroskopie atomárních sil
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PŘIBYL Jan PEŠL Martin AĆIMOVIĆ Ivana VILOTIĆ Aleksandra MELI Albano ROTREKL Vladimír DVOŘÁK Petr SKLÁDAL Petr

Rok publikování 2014
Druh Vyžádané přednášky
Citace
Popis Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) can be differentiated into functional cardiomyocytes (CMs) in vitro serving as an important source of cells for regenerative medicine and modeling of hart diseases. Differentiation of hESCs and hiPSCs to the cardiac lineage can be done using different methods as well as different cultivation matrices. These factors can dramatically influence the differentiation outcome. The resulting cardiomyocytes differ not only in yield but also by their biomechanical properties which can be considered as one of the key parameters of the cells quality. Robust and reliable method to assess the biomechanical properties of cardiomyocytes can be provided by Atomic Force Microscopy. It is part of scanning probe microscopy, originally developed for visualization of solid-phase immobilized samples with a high resolution. Interaction of cantileverbound tip with rigid sample is evaluated in order to obtain surface properties (profile view, phase imaging, etc.). In this study, interaction of cantilever with beating cell cluster was evaluated, and real-time recorded deflection vs. time curves were further evaluated to get information about the force and frequency of the biological movement. We implemented the AFM microscope in the apparatus for monitoring of biomechanical properties of cardiomyocytes. Tip located on the cardiomyocyte surface mediates cantilever deflection in real time caused by cell movement (beating). Force of beating can be measured in this way, when cantilever stiffness is multiplied by its deflection determined by AFM measurement. AFM based method is the only one able to routinely monitor beating force of cardiomyocytes in absolute values. Force values in units/tens of nN and beating rate proceeding in range 50–90 bpm were usually measured. Experimental setup, data evaluation process and examples of effect of various drugs affecting biomechanical properties of cell clusters will be presented. Acknowledgement The work has been supported by CEITEC – Central European Institute of Technology (CZ.1.05/1.1.00/ 02.0068)and FNUSA-ICRC (no. CZ.1.05/1.1.00/02. 0123) from the European Regional Development Fund
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