{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T03:53:01Z","timestamp":1791431581725,"version":"4.3.3"},"reference-count":35,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T00:00:00Z","timestamp":1773360000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Biosci."],"abstract":"<jats:p>\n                    Human peripheral blood mononuclear cells (PBMCs) can be easily sampled from healthy individuals and patients. Density gradient isolation from human blood or leukocyte concentrates yields a mononuclear cell population of mainly lymphocytes, monocytes, and natural killer (NK) cells. PBMCs are vital circulating cells of the immune system and rely on oxidative phosphorylation (OXPHOS) for their energy production. OXPHOS capacity can be assessed using oxygraphy in intact and permeabilized PBMCs and has been used to investigate disorders of the immune system, but also, similarly to platelets, employed as a bioenergetic biomarker, that is, a \u201cliquid biopsy\u201d of disease conditions unrelated to immune dysregulation. Here, we present some key aspects of mitochondrial respiration in PBMCs isolated from leukocyte concentrates and whole blood using the Oroboros O2k oxygraph. We assessed the limits of sample amount and the impact of storage time and temperature and explored critical aspects of digitonin permeabilization. Furthermore, we provide respiratory rates and internal ratios from healthy controls using simple and comprehensive protocols for intact and permeabilized PBMCs, respectively. We conclude that detailed information on OXPHOS capacity in PBMCs can be reproducibly assessed\n                    <jats:italic>ex vivo<\/jats:italic>\n                    , but that great care must be taken during permeabilization to achieve correct measures of respiratory rates.\n                  <\/jats:p>","DOI":"10.3389\/fmolb.2026.1756657","type":"journal-article","created":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T05:26:36Z","timestamp":1773379596000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Mitochondrial respiration in human peripheral blood mononuclear cells: methodology and influence of permeabilization and storage"],"prefix":"10.3389","volume":"13","author":[{"given":"Shusuke","family":"Sekine","sequence":"first","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]},{"name":"Department of Anesthesiology, Tokyo Medical University","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Imen","family":"Chamkha","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evelina","family":"Elm\u00e9r","sequence":"additional","affiliation":[{"name":"Division of Transfusion Medicine, Department of Laboratory Medicine, Lund University","place":["Lund, Sweden"]},{"name":"Department of Clinical Immunology and Transfusion Medicine, Sk\u00e5ne University Hospital","place":["Lund, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eleonor","family":"\u00c5sander Frostner","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emil","family":"Westerlund","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]},{"name":"Emergency Department, Kung\u00e4lv Hospital","place":["Kung\u00e4lv, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianshi","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes","family":"Ehinger","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]},{"name":"Department of Clinical Sciences Lund, Otorhinolaryngology, Head and Neck Surgery, Lund University","place":["Lund, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fredrik","family":"Sj\u00f6vall","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]},{"name":"Department of Intensive- and perioperative Care, Sk\u00e5ne University Hospital","place":["Malm\u00f6, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroyuki","family":"Uchino","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Tokyo Medical University","place":["Tokyo, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eskil","family":"Elm\u00e9r","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences Lund, Mitochondrial Medicine, Lund University","place":["Lund, Sweden"]},{"name":"Clinical Neurophysiology, Medical Imaging and Physiology, Sk\u00e5ne University Hospital","place":["Lund, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,3,13]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"101890","DOI":"10.1016\/j.mito.2024.101890","article-title":"Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis","volume":"77","author":"Bager Christensen","year":"2024","journal-title":"Mitochondrion"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.26124\/bec.2024-0008","article-title":"Stability of mitochondrial respiration medium used in high-resolution respirometry with living and permeabilized cells","author":"Baglivo","year":"2024","journal-title":"Bioenerg. 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