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Two enzymatic approaches, collagenase A and dispase, are widely employed for endothelial cell isolation. In this study, the utility of both enzymatic approaches, alone and in combination, for endothelial cell isolation from juvenile and adult mouse lungs was assessed, considering the number, viability, and subtype composition of recovered endothelial cell pools. Collagenase A yielded an 8\u201012\u2010fold superior recovery of viable endothelial cells from lung tissue from developing mouse pups, compared to dispase, although dispase proved superior in efficiency for epithelial cell recovery. Single\u2010cell RNA\u2010Seq revealed that the collagenase A approach yielded a diverse endothelial cell subtype composition of recovered endothelial cell pools, with broad representation of arterial, capillary, venous, and lymphatic lung endothelial cells; while the dispase approach yielded a recovered endothelial cell pool highly enriched for one subset of general capillary endothelial cells, but poor representation of other endothelial cells subtypes. These data indicate that tissue dissociation markedly influences the recovery of endothelial cells, and the endothelial subtype composition of recovered endothelial cell pools, as assessed by single\u2010cell RNA\u2010Seq.<\/jats:p>","DOI":"10.1002\/cyto.a.24843","type":"journal-article","created":{"date-parts":[[2024,4,26]],"date-time":"2024-04-26T09:23:41Z","timestamp":1714123421000},"page":"521-535","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Impact of different tissue dissociation protocols on endothelial cell recovery from developing mouse lungs"],"prefix":"10.1002","volume":"105","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5352-6773","authenticated-orcid":false,"given":"Francesco","family":"Palumbo","sequence":"first","affiliation":[{"name":"Flow Cytometry Core Facility, School of Life Sciences \u00c9cole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)  Lausanne 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Liebig University Giessen  Giessen Germany"},{"name":"Department of Translational Pulmonology and the Translational Lung Research Center Heidelberg (TLRC) Heidelberg University Hospital, member of the German Center for Lung Research (DZL)  Heidelberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9906-1628","authenticated-orcid":false,"given":"Patty J.","family":"Lee","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care and Sleep Medicine Icahn School of Medicine at Mount Sinai  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5594-4549","authenticated-orcid":false,"given":"Stefan","family":"G\u00fcnther","sequence":"additional","affiliation":[{"name":"Deep Sequencing Platform Max Planck Institute for Heart and Lung Research, Bad Nauheim, member of the German Center for Lung Research (DZL)  Bad Nauheim 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