Dual cardiomyocyte cluster detection the different types of arrhythmogenic potential of caffein

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PEŠL Martin PIVATO Roberto KLIMOVIČ Šimon KABANOV Daniil SVĚRÁK Filip PŘIBYL Jan ROTREKL Vladimír

Rok publikování 2022
Druh Článek v odborném periodiku
Časopis / Zdroj AFM BioMed Conference 2022 Abstract Book Nagoya - Okazaki
Fakulta / Pracoviště MU

Lékařská fakulta

Citace
www https://afmbiomed.org/scientific-program-3.aspx
Klíčová slova Cell-based biosensor; Atomic force microscopy; Human embryonic stem cells; Cardiac arrhythmia; Cardiomyocytes; Caffeine
Přiložené soubory
Popis Single cardiomyocytes present unstable beating patterns, which can be partially compensated in a clustered syncytium. Here we present a novel biosensor composed of two clusters of cardiomyocytes serving for advanced detection of cardiac arrhythmias and subsequent drug testing. hESC's clusters were differentiated into CMs syncytium. Two such clusters were seeded on adherent plates in the vicinity and spontaneously formed electrically connected syncytium resembling the human myocardium and its conductive system. Contraction of twin clustered spontaneously synchronized within 4 days. The Atomic Force Microscope (AFM) vertical deflection enabled measurement and calculation of absolute cardiomyocyte contraction force [1], while lateral force measurement can distinguish synchronized and independent contraction behavior of twin clusters [2]. Caffeine induces arrhythmia as a model drug [3]. Subsequent beating patterns were monitored by AFM lateral force recording and calcium fluorescence imaging as a reference method for describing non-synchronized contractions of cardiomyocytes. Caffeine addition increased the beat rate of the syncytium. This was reflected by both the change in vertical and lateral deflection. A significant increase in standard deviation was observed immediately after caffeine injection for both the lateral and the vertical deflection. Further, caffeine affected the synchronization of vertical and lateral displacement of the AFM cantilever. The lateral and vertical deflection peaks followed each other closely, adding caffeine produced independent lateral and vertical deflections, interpreted as defects in signal spreading through the bridge resulting in the irregular beat of the two clusters. References [1] Pesl M., Pribyl J., Acimovic A. et al. Biosens. Bioelectron. 85, 2016,10.1016/j.bios.2016.05.073 [2] Pivato R, Klimovic S, Kabanov D, et al Anal Chim Acta. 2022 Jul 11;1216:339959. doi: 10.1016/j.aca.2022.339959. [3] Voskoboinik A., Kalman J.M., Kistler P.M. JACC (J. Am. Coll. Cardiol.): Clinic. Electrophys., 4 (2018), pp. 425-432, 10.1016/j.jacep.2018.01.012 Acknowledgments Ministry of Health of the Czech Republic, grant NU20-06-00156 has financially supported this work. CIISB, Instruct-CZ Centre of Instruct-ERIC EU consortium, funded by MEYS CR infrastructure project LM2018127, is gratefully acknowledged for the financial support of the measurements at the CF Nanobiotechnology.
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