{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:26:02Z","timestamp":1760235962310,"version":"build-2065373602"},"reference-count":14,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,10,16]],"date-time":"2021-10-16T00:00:00Z","timestamp":1634342400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the brain foundation","award":["no number"],"award-info":[{"award-number":["no number"]}]},{"DOI":"10.13039\/501100006065","name":"Government of Western Australia Department of Health","doi-asserted-by":"publisher","award":["biobank2021"],"award-info":[{"award-number":["biobank2021"]}],"id":[{"id":"10.13039\/501100006065","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001799","name":"Murdoch University","doi-asserted-by":"publisher","award":["Research Training Programme Scholarship"],"award-info":[{"award-number":["Research Training Programme Scholarship"]}],"id":[{"id":"10.13039\/501100001799","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100019111","name":"PERRON INSTITUTE FOR NEUROLOGICAL AND TRANSLATIONAL SCIENCE","doi-asserted-by":"publisher","award":["Byron Kakulas top-up scholarship"],"award-info":[{"award-number":["Byron Kakulas top-up scholarship"]}],"id":[{"id":"10.13039\/501100019111","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["MPs"],"abstract":"<jats:p>In inflammatory myopathies, the self-reactive immune cells involved in muscle aggression have been studied mostly using histological assessment of muscle biopsy sections; this methodology provides the advantage of visualizing and identifying cells within the tissue, but it does not allow further investigation. To gain access to live and isolated cells, many studies utilized blood samples; however, in the absence of biological tools to discriminate the leukocytes associated with the autoimmune process from those that emerged from responses against pathogens, the information observed on circulating immune cells often lacks in specificity, and thus result interpretation may prove difficult. In order to selectively retrieve self-reactive immune cells, we developed a protocol to isolate live leukocytes from human muscle biopsies, which allows for further analysis using a large range of methodologies. The protocol uses enzymatic digestion to release live leukocytes from freshly collected skeletal muscle samples, followed by filtration and separation of the leukocytes from the myocytes by density gradient centrifugation. The isolated cells can be submitted immediately to various analysis strategies to characterize ex vivo the specific cellular and molecular mechanisms responsible for self-directed immune muscle aggression or may be placed in culture for expansion.<\/jats:p>","DOI":"10.3390\/mps4040075","type":"journal-article","created":{"date-parts":[[2021,10,18]],"date-time":"2021-10-18T13:59:52Z","timestamp":1634565592000},"page":"75","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Isolation of Live Leukocytes from Human Inflammatory Muscles"],"prefix":"10.3390","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6081-8494","authenticated-orcid":false,"given":"Jerome D.","family":"Coudert","sequence":"first","affiliation":[{"name":"Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Murdoch, WA 6150, Australia"},{"name":"Perron Institute for Neurological and Translational Science, Nedlands, WA 6009, Australia"},{"name":"School of Medicine, University of Notre Dame, Fremantle, WA 6160, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emily","family":"McLeish","sequence":"additional","affiliation":[{"name":"Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Murdoch, WA 6150, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anuradha","family":"Sooda","sequence":"additional","affiliation":[{"name":"Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Murdoch, WA 6150, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nataliya","family":"Slater","sequence":"additional","affiliation":[{"name":"Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Murdoch, WA 6150, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kelly","family":"Beer","sequence":"additional","affiliation":[{"name":"Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Murdoch, WA 6150, Australia"},{"name":"Perron Institute for Neurological and Translational Science, Nedlands, WA 6009, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shereen","family":"Paramalingam","sequence":"additional","affiliation":[{"name":"Department of Rheumatology, Fiona Stanley Hospital, Murdoch, WA 6150, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Phillipa J.","family":"Lamont","sequence":"additional","affiliation":[{"name":"Neurogenetic Unit, Royal Perth Hospital, Perth, WA 6000, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Merrilee","family":"Needham","sequence":"additional","affiliation":[{"name":"Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Murdoch, WA 6150, Australia"},{"name":"School of Medicine, University of Notre Dame, Fremantle, WA 6160, Australia"},{"name":"Department of Neurology, Fiona Stanley Hospital, Murdoch, WA 6150, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1734","DOI":"10.1056\/NEJMra1402225","article-title":"Inflammatory muscle diseases","volume":"372","author":"Dalakas","year":"2015","journal-title":"N. 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