J 2019

Simultaneous study of mechanobiology and calcium dynamics on hESC-derived cardiomyocytes clusters

CALUORI, Guido, Jan PŘIBYL, Vratislav CMIEL, Martin PEŠL, Tomas POTOCNAK et. al.

Basic information

Original name

Simultaneous study of mechanobiology and calcium dynamics on hESC-derived cardiomyocytes clusters

Authors

CALUORI, Guido (380 Italy, belonging to the institution), Jan PŘIBYL (203 Czech Republic, guarantor, belonging to the institution), Vratislav CMIEL (203 Czech Republic), Martin PEŠL (203 Czech Republic, belonging to the institution), Tomas POTOCNAK (203 Czech Republic), Ivo PROVAZNÍK (203 Czech Republic), Petr SKLÁDAL (203 Czech Republic, belonging to the institution) and Vladimír ROTREKL (203 Czech Republic, belonging to the institution)

Edition

Journal of Molecular Recognition, Hoboken, Wiley, 2019, 0952-3499

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 2.214

RIV identification code

RIV/00216224:14740/19:00107155

Organization unit

Central European Institute of Technology

UT WoS

000459589700001

Keywords in English

calcium imaging; atomic force microscopy; human stem cell-derived cardiomyocytes; in vitro models; fluorescence microscopy; cardiac differentiation; caffeine; embryoid bodies; biosignals filtering

Tags

International impact, Reviewed
Změněno: 24/10/2024 11:51, Mgr. Adéla Pešková

Abstract

V originále

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.

Links

GA18-24089S, research and development project
Name: Kvantitativní fázová mikroskopie pro 3D kvalitativní charakterizaci nádorových buněk
Investor: Czech Science Foundation
LM2015043, research and development project
Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
LQ1601, research and development project
Name: CEITEC 2020 (Acronym: CEITEC2020)
Investor: Ministry of Education, Youth and Sports of the CR
MUNI/A/1087/2018, interní kód MU
Name: Molekulární a buněčná biologie pro biomedicínské vědy
Investor: Masaryk University, Category A