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The presumption that cardiomyocytes must be isolated immediately after heart procurement to avoid deterioration implies that transgenic mouse lines must be present on site, causes schedule inflexibility, and hampers collaborations, thereby increasing the number, suffering, and costs of animals. This study challenges this presumption by investigating whether the cell isolation can be postponed for 24\u00a0h without affecting the results. Adult mouse hearts were subjected to enzymatic myocyte isolation immediately after excision (CTRL) or after 24\u00a0h of cold storage (CS). Sufficient numbers of viable cardiomyocytes were obtained in all groups. The transverse-axial tubular system was unchanged in CS versus CTRL. No significant changes were detected in cell capacitance, resting membrane potential, action potential shape and duration, amplitudes, and kinetics of the K<jats:sup>+<\/jats:sup> currents I<jats:sub>to<\/jats:sub>, I<jats:sub>K1<\/jats:sub>, and I<jats:sub>K<\/jats:sub>. Sarcomere length, contractility, and relaxation as well as Ca<jats:sup>2+<\/jats:sup> signals were equivalent in CS and CTRL at pacing rates of 1\u20134\u00a0Hz. Mitochondrial function assays also yielded equivalence. RNA sequencing yielded only 128 differentially expressed genes, which were mainly related to immune cell function and inflammation. Key findings were reproduced in infarcted mouse hearts, which were shipped overnight as a proof of principle. This study demonstrates that the isolation of cardiomyocytes can be postponed up to 24\u00a0h after the procurement of the heart. This opens up new possibilities for collaboration between different laboratories, increases experimental flexibility, and allows to reduce the number of experimental animals by avoiding unnecessary propagation of transgenic lines.<\/jats:p>","DOI":"10.1007\/s00395-025-01131-y","type":"journal-article","created":{"date-parts":[[2025,7,29]],"date-time":"2025-07-29T08:32:37Z","timestamp":1753777957000},"page":"1055-1074","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Cold storage of mouse hearts prior to cardiomyocyte isolation preserves electromechanical function, microstructure, and gene expression for 24\u00a0h"],"prefix":"10.1007","volume":"120","author":[{"given":"Benedikt","family":"Pfeilschifter","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aiora","family":"Martinez-Vilchez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zafar","family":"Iqbal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6356-735X","authenticated-orcid":false,"given":"Prapassorn","family":"Potue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominik J.","family":"Fiegle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karoline","family":"Morhenn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander P.","family":"Schwoerer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tilmann","family":"Volk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3127-1693","authenticated-orcid":false,"given":"Thomas","family":"Seidel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,29]]},"reference":[{"key":"1131_CR1","doi-asserted-by":"publisher","first-page":"370","DOI":"10.1113\/expphysiol.2007.040659","volume":"93","author":"T Banyasz","year":"2008","unstructured":"Banyasz T, Lozinskiy I, Payne CE, Edelmann S, Norton B, Chen B, Chen-Izu Y, Izu LT, Balke CW (2008) Transformation of adult rat cardiac myocytes in primary culture. 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