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Preserving RBCs for clinical transfusion remains a challenge due to storage\u2010induced damage and limitations of traditional freezing methods. This study investigates isochoric freezing\u2014a constant\u2010volume, high\u2010pressure cryopreservation technique that suppresses ice formation\u2014as an alternative approach for RBC cryopreservation. RBC samples were preserved under isochoric freezing conditions at \u22122.5\u00b0C to \u221215\u00b0C with corresponding pressures of 31\u2013156\u2009MPa and a comparator supercooled control group. Hemolysis, cell count, morphology, and membrane integrity were assessed using hematological analyses, imaging flow cytometry, and dextran permeability assays. It is reported that hemolysis and morphological deterioration increased with decreasing temperature and rising pressure, with higher membrane damage compared to supercooling. Lower\u2010temperature isochoric freezing resulted in loss of membrane integrity that was irreversible. While isochoric freezing minimized ice formation, elevated pressures adversely affected RBC viability. These findings highlight critical pressure\u2010temperature thresholds necessary for optimizing isochoric freezing protocols for RBC preservation and inform future development of safer, long\u2010term blood storage strategies.<\/jats:p>","DOI":"10.1002\/bit.70145","type":"journal-article","created":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T06:42:09Z","timestamp":1767595329000},"page":"973-982","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Evaluating Isochoric Freezing as a Strategy for Storage of Red Blood Cells"],"prefix":"10.1002","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9459-641X","authenticated-orcid":false,"given":"Yuanheng","family":"Zhao","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine and Pathology University of Alberta Edmonton Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaitlyn","family":"Wingnean","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology University of Alberta Edmonton Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nishaka","family":"William","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology University of Alberta Edmonton Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1445-827X","authenticated-orcid":false,"given":"Jason P.","family":"Acker","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology University of Alberta Edmonton Alberta Canada"},{"name":"Innovation and Portfolio Management, Canadian Blood Services Edmonton Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,1,4]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41538-025-00542-4"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ifset.2018.10.016"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1080\/10498850.2020.1785602"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1749-6632.2009.05182.x"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/trf.17471"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1115\/1.4054217"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1039\/d3fb00146f"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-021-00549-7"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.23987"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2001227117"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-26309-5"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/0140-7007(80)90045-6"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.IJREFRIG.2023.10.014"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/vox.12218"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-05636-0"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-024-06463-4"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.JTHERBIO.2022.103196"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI91309"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1364\/boe.9.002755"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.JTHERBIO.2020.102689"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/trf.16017"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-025-18028-4"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12393-021-09284-x"},{"key":"e_1_2_9_25_1","unstructured":"Organization W. 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