{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T18:53:54Z","timestamp":1783536834003,"version":"3.55.0"},"reference-count":34,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,1,11]],"date-time":"2022-01-11T00:00:00Z","timestamp":1641859200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["396913060"],"award-info":[{"award-number":["396913060"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["422681845"],"award-info":[{"award-number":["422681845"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000780","name":"European Union","doi-asserted-by":"publisher","award":["860974"],"award-info":[{"award-number":["860974"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["Cells"],"abstract":"<jats:p>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\u20138 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.<\/jats:p>","DOI":"10.3390\/cells11020233","type":"journal-article","created":{"date-parts":[[2022,1,11]],"date-time":"2022-01-11T14:05:07Z","timestamp":1641909907000},"page":"233","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost\u2013Benefit Balance of Cardiac Primary Cell Research"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5454-2962","authenticated-orcid":false,"given":"Joachim","family":"Greiner","sequence":"first","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teresa","family":"Schiatti","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenzel","family":"Kaltenbacher","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marica","family":"Dente","sequence":"additional","affiliation":[{"name":"Department of Experimental and Clinical Medicine, Division of Physiology, University of Florence, 50134 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0448-8643","authenticated-orcid":false,"given":"Alina","family":"Semenjakin","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8772-3924","authenticated-orcid":false,"given":"Thomas","family":"Kok","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominik","family":"Fiegle","sequence":"additional","affiliation":[{"name":"Institute of Cellular and Molecular Physiology, Friedrich-Alexander-University of Erlangen-N\u00fcrnberg, 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3127-1693","authenticated-orcid":false,"given":"Thomas","family":"Seidel","sequence":"additional","affiliation":[{"name":"Institute of Cellular and Molecular Physiology, Friedrich-Alexander-University of Erlangen-N\u00fcrnberg, 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3813-3127","authenticated-orcid":false,"given":"Ursula","family":"Ravens","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0416-6270","authenticated-orcid":false,"given":"Peter","family":"Kohl","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"},{"name":"CIBSS Centre for Integrative Biological Signalling Studies, University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R\u00e9mi","family":"Peyronnet","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Rog-Zielinska","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg\u2014Bad Krozingen and Faculty of Medicine, Albert-Ludwig University of Freiburg, 79110 Freiburg im Breisgau, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,11]]},"reference":[{"key":"ref_1","first-page":"16","article-title":"Intact isolated muscle cells from the adult rat heart","volume":"225","author":"Gould","year":"1972","journal-title":"J. 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