{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T01:26:30Z","timestamp":1793928390982,"version":"4.8.0"},"reference-count":33,"publisher":"American Physiological Society","issue":"2","funder":[{"name":"IHU OPERA","award":["ANR-10-IBHU-004"],"award-info":[{"award-number":["ANR-10-IBHU-004"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["American Journal of Physiology-Cell Physiology"],"published-print":{"date-parts":[[2020,2,1]]},"abstract":"<jats:p> Cardiovascular diseases remain the leading cause of death worldwide. Although major therapeutic progress has been made during the past decades, a better understanding of the underlying mechanisms will certainly help to improve patient\u2019s prognosis. In vitro models, particularly adult mouse cardiomyocytes, have been largely used; however, their fragility and large size are major obstacles to the use of flow cytometry. Conventional techniques, such as cell imaging, require the use of large numbers of animals and are time consuming. Here, we described a new, simple, and rapid one-day protocol using living adult mouse cardiomyocytes in suspension exposed to hypoxia-reoxygenation that allows a multilabeling analysis by flow cytometry. Several parameters can be measured by fluorescent probes labeling to assess cell viability (propidium iodide, calcein-AM, and Sytox Green), mitochondrial membrane potential [DilC1(5) and TMRM], reactive oxygen species production (MitoSOX Red), and mitochondrial mass (MitoTracker Deep Red). We address the robustness and sensitivity of our model using a cardioprotective agent, cyclosporine A. Overall, our new experimental set-up offers a high-speed quantitative multilabeling analysis of adult mouse cardiomyocytes exposed to hypoxia-reoxygenation. Our model might be interesting to investigate other cellular stresses (oxidative and inflammation) or to perform pharmacological screening. <\/jats:p>","DOI":"10.1152\/ajpcell.00393.2019","type":"journal-article","created":{"date-parts":[[2019,12,26]],"date-time":"2019-12-26T04:34:13Z","timestamp":1577334853000},"page":"C439-C447","source":"Crossref","is-referenced-by-count":8,"title":["An innovative sequence of hypoxia-reoxygenation on adult mouse cardiomyocytes in suspension to perform multilabeling analysis by flow cytometry"],"prefix":"10.1152","volume":"318","author":[{"given":"Alexandre","family":"Paccalet","sequence":"first","affiliation":[{"name":"Universit\u00e9 Lyon, CarMeN Laboratory, Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale, Institut National de la Recherche Agronomique, Institut National des Sciences Appliqu\u00e9es de Lyon, Lyon, Universit\u00e9 Claude Bernard Lyon 1, Bron, 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