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Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research

Freshly isolated primary cardiomyocytes (CM) are indispensable for cardiac research. Experimental CM research is generally incompatible with life of the donor animal, while human heart samples are usually small and scarce. CM isolation from animal hearts, traditionally performed by coronary artery p...

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Autores principales: Greiner, Joachim, Schiatti, Teresa, Kaltenbacher, Wenzel, Dente, Marica, Semenjakin, Alina, Kok, Thomas, Fiegle, Dominik J., Seidel, Thomas, Ravens, Ursula, Kohl, Peter, Peyronnet, Rémi, Rog-Zielinska, Eva A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8773758/
https://www.ncbi.nlm.nih.gov/pubmed/35053351
http://dx.doi.org/10.3390/cells11020233
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author Greiner, Joachim
Schiatti, Teresa
Kaltenbacher, Wenzel
Dente, Marica
Semenjakin, Alina
Kok, Thomas
Fiegle, Dominik J.
Seidel, Thomas
Ravens, Ursula
Kohl, Peter
Peyronnet, Rémi
Rog-Zielinska, Eva A.
author_facet Greiner, Joachim
Schiatti, Teresa
Kaltenbacher, Wenzel
Dente, Marica
Semenjakin, Alina
Kok, Thomas
Fiegle, Dominik J.
Seidel, Thomas
Ravens, Ursula
Kohl, Peter
Peyronnet, Rémi
Rog-Zielinska, Eva A.
author_sort Greiner, Joachim
collection PubMed
description Freshly isolated primary cardiomyocytes (CM) are indispensable for cardiac research. Experimental CM research is generally incompatible with life of the donor animal, while human heart samples are usually small and scarce. CM isolation from animal hearts, traditionally performed by coronary artery perfusion of enzymes, liberates millions of cells from the heart. However, due to progressive cell remodeling following isolation, freshly isolated primary CM need to be used within 4–8 h post-isolation for most functional assays, meaning that the majority of cells is essentially wasted. In addition, coronary perfusion-based isolation cannot easily be applied to human tissue biopsies, and it does not straightforwardly allow for assessment of regional differences in CM function within the same heart. Here, we provide a method of multi-day CM isolation from one animal heart, yielding calcium-tolerant ventricular and atrial CM. This is based on cell isolation from cardiac tissue slices following repeated (usually overnight) storage of the tissue under conditions that prolong CM viability beyond the day of organ excision by two additional days. The maintenance of cells in their near-native microenvironment slows the otherwise rapid structural and functional decline seen in isolated CM during attempts for prolonged storage or culture. Multi-day slice-based CM isolation increases the amount of useful information gained per animal heart, improving reproducibility and reducing the number of experimental animals required in basic cardiac research. It also opens the doors to novel experimental designs, including exploring same-heart regional differences.
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spelling pubmed-87737582022-01-21 Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research Greiner, Joachim Schiatti, Teresa Kaltenbacher, Wenzel Dente, Marica Semenjakin, Alina Kok, Thomas Fiegle, Dominik J. Seidel, Thomas Ravens, Ursula Kohl, Peter Peyronnet, Rémi Rog-Zielinska, Eva A. Cells Article Freshly isolated primary cardiomyocytes (CM) are indispensable for cardiac research. Experimental CM research is generally incompatible with life of the donor animal, while human heart samples are usually small and scarce. CM isolation from animal hearts, traditionally performed by coronary artery perfusion of enzymes, liberates millions of cells from the heart. However, due to progressive cell remodeling following isolation, freshly isolated primary CM need to be used within 4–8 h post-isolation for most functional assays, meaning that the majority of cells is essentially wasted. In addition, coronary perfusion-based isolation cannot easily be applied to human tissue biopsies, and it does not straightforwardly allow for assessment of regional differences in CM function within the same heart. Here, we provide a method of multi-day CM isolation from one animal heart, yielding calcium-tolerant ventricular and atrial CM. This is based on cell isolation from cardiac tissue slices following repeated (usually overnight) storage of the tissue under conditions that prolong CM viability beyond the day of organ excision by two additional days. The maintenance of cells in their near-native microenvironment slows the otherwise rapid structural and functional decline seen in isolated CM during attempts for prolonged storage or culture. Multi-day slice-based CM isolation increases the amount of useful information gained per animal heart, improving reproducibility and reducing the number of experimental animals required in basic cardiac research. It also opens the doors to novel experimental designs, including exploring same-heart regional differences. MDPI 2022-01-11 /pmc/articles/PMC8773758/ /pubmed/35053351 http://dx.doi.org/10.3390/cells11020233 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Greiner, Joachim
Schiatti, Teresa
Kaltenbacher, Wenzel
Dente, Marica
Semenjakin, Alina
Kok, Thomas
Fiegle, Dominik J.
Seidel, Thomas
Ravens, Ursula
Kohl, Peter
Peyronnet, Rémi
Rog-Zielinska, Eva A.
Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research
title Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research
title_full Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research
title_fullStr Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research
title_full_unstemmed Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research
title_short Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research
title_sort consecutive-day ventricular and atrial cardiomyocyte isolations from the same heart: shifting the cost–benefit balance of cardiac primary cell research
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8773758/
https://www.ncbi.nlm.nih.gov/pubmed/35053351
http://dx.doi.org/10.3390/cells11020233
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