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Article

Live-Cell Imaging of the Contractile Velocity and Transient Intracellular Ca2+ Fluctuations in Human Stem Cell-Derived Cardiomyocytes

by
Aviseka Acharya
1,
Harshal Nemade
1,
Krishna Rajendra Prasad
1,
Khadija Khan
1,
Jürgen Hescheler
1,
Nick Blackburn
2,
Ruth Hemmersbach
3,
Symeon Papadopoulos
1 and
Agapios Sachinidis
1,4,*
1
Working Group Sachinidis, Center for Physiology, Faculty of Medicine and University Hospital Cologne, The University of Cologne, 50931 Cologne, Germany
2
Bioras Company, Kaarsbergsvej 2, 8400 Ebeltoft, Denmark
3
German Aerospace Center, Institute of Aerospace Medicine, Gravitational Biology, Linder Hoehe, 51147 Cologne, Germany
4
Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany
*
Author to whom correspondence should be addressed.
Cells 2022, 11(8), 1280; https://doi.org/10.3390/cells11081280
Submission received: 4 March 2022 / Revised: 25 March 2022 / Accepted: 6 April 2022 / Published: 9 April 2022
(This article belongs to the Collection Applications of Stem Cells in Cardiovascular Functional Genomics 2022)
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Abstract

Live-cell imaging techniques are essential for acquiring vital physiological and pathophysiological knowledge to understand and treat heart disease. For live-cell imaging of transient alterations of [Ca2+]i in human cardiomyocytes, we engineered human-induced pluripotent stem cells carrying a genetically-encoded Ca2+-indicator (GECI). To monitor sarcomere shortening and relaxation in cardiomyocytes in real-time, we generated a α-cardiac actinin (ACTN2)-copepod (cop) green fluorescent protein (GFP+)-human-induced pluripotent stem cell line by using the CRISPR-Cas9 and a homology directed recombination approach. The engineered human-induced pluripotent stem cells were differentiated in transgenic GECI-enhanced GFP+-cardiomyocytes and ACTN2-copGFP+-cardiomyocytes, allowing real-time imaging of [Ca2+]i transients and live recordings of the sarcomere shortening velocity of ACTN2-copGFP+-cardiomyocytes. We developed a video analysis software tool to quantify various parameters of sarcoplasmic Ca2+ fluctuations recorded during contraction of cardiomyocytes and to calculate the contraction velocity of cardiomyocytes in the presence and absence of different drugs affecting cardiac function. Our cellular and software tool not only proved the positive and negative inotropic and lusitropic effects of the tested cardioactive drugs but also quantified the expected effects precisely. Our platform will offer a human-relevant in vitro alternative for high-throughput drug screenings, as well as a model to explore the underlying mechanisms of cardiac diseases.
Keywords: hiPSCs; contractile velocity of cardiomyocytes; CRISPR-Cas9; genetically encoded Ca2+-indicator; drug screening hiPSCs; contractile velocity of cardiomyocytes; CRISPR-Cas9; genetically encoded Ca2+-indicator; drug screening
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MDPI and ACS Style

Acharya, A.; Nemade, H.; Rajendra Prasad, K.; Khan, K.; Hescheler, J.; Blackburn, N.; Hemmersbach, R.; Papadopoulos, S.; Sachinidis, A. Live-Cell Imaging of the Contractile Velocity and Transient Intracellular Ca2+ Fluctuations in Human Stem Cell-Derived Cardiomyocytes. Cells 2022, 11, 1280. https://doi.org/10.3390/cells11081280

AMA Style

Acharya A, Nemade H, Rajendra Prasad K, Khan K, Hescheler J, Blackburn N, Hemmersbach R, Papadopoulos S, Sachinidis A. Live-Cell Imaging of the Contractile Velocity and Transient Intracellular Ca2+ Fluctuations in Human Stem Cell-Derived Cardiomyocytes. Cells. 2022; 11(8):1280. https://doi.org/10.3390/cells11081280

Chicago/Turabian Style

Acharya, Aviseka, Harshal Nemade, Krishna Rajendra Prasad, Khadija Khan, Jürgen Hescheler, Nick Blackburn, Ruth Hemmersbach, Symeon Papadopoulos, and Agapios Sachinidis. 2022. "Live-Cell Imaging of the Contractile Velocity and Transient Intracellular Ca2+ Fluctuations in Human Stem Cell-Derived Cardiomyocytes" Cells 11, no. 8: 1280. https://doi.org/10.3390/cells11081280

APA Style

Acharya, A., Nemade, H., Rajendra Prasad, K., Khan, K., Hescheler, J., Blackburn, N., Hemmersbach, R., Papadopoulos, S., & Sachinidis, A. (2022). Live-Cell Imaging of the Contractile Velocity and Transient Intracellular Ca2+ Fluctuations in Human Stem Cell-Derived Cardiomyocytes. Cells, 11(8), 1280. https://doi.org/10.3390/cells11081280

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