Supercooled storage of red blood cells slows down the metabolic storage lesion

Supercooled storage of red blood cells slows down the metabolic storage lesion | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Supercooled storage of red blood cells slows down the metabolic storage lesion Travis Nemkov, Ziya Isiksacan, Nishaka William, Rahime Senturk, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5256734/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 03 Oct, 2025 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract Red blood cell (RBC) transfusion, a life-saving intervention, is limited by reduced RBC potency over time. Cold storage at + 4°C for up to 42 days can reduce transfusion efficacy due to alterations termed the "storage lesion." Strategies to mitigate the storage lesion include alkaline additive solutions and supercooled storage to extend storage by reducing metabolic stresses. However, RBC metabolism during supercooled storage in standard or alkaline additives remains unstudied. This study, thus, investigated the impact of storage additives (alkaline E-Sol5 and standard SAGM) and temperatures (+ 4°C, -4°C, -8°C) on RBC metabolism during 21- and 42-days storage using high-throughput metabolomics. RBCs stored with E-Sol5 showed increased glycolysis and higher ratios of reduced to oxidized glutathione compared to SAGM. Supercooled storage at -4°C showed markedly lower hemolysis than − 8°C, preserved adenylate pools, decreased glucose consumption, and reduced lactate accumulation and pentose phosphate pathway activation. The combination of supercooled storage and E-Sol5 helped to preserve ATP and 2,3-DPG reservoirs, while preventing catabolism and free fatty acid accumulation. While supercooled storage with E-Sol5 offers a promising alternative to standard storage, preserving RBC metabolic and functional quality, further research is necessary to validate and improve on these foundational findings.​​ Biological sciences/Biotechnology/Metabolomics Biological sciences/Biotechnology Health sciences/Medical research Physical sciences/Engineering Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 03 Oct, 2025 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 26 May, 2025 Reviews received at journal 23 May, 2025 Reviewers agreed at journal 07 May, 2025 Reviews received at journal 26 Jan, 2025 Reviewers agreed at journal 17 Jan, 2025 Reviewers invited by journal 10 Nov, 2024 Editor assigned by journal 10 Nov, 2024 Editor invited by journal 30 Oct, 2024 Submission checks completed at journal 28 Oct, 2024 First submitted to journal 13 Oct, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5256734","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":407320362,"identity":"73b1a6a0-7f82-46af-8b11-60069eb8cc18","order_by":0,"name":"Travis Nemkov","email":"","orcid":"","institution":"University of Colorado Anschutz Medical Campus","correspondingAuthor":false,"prefix":"","firstName":"Travis","middleName":"","lastName":"Nemkov","suffix":""},{"id":407320363,"identity":"6cdf1799-9133-4f62-b0da-63c16f7e7fca","order_by":1,"name":"Ziya Isiksacan","email":"","orcid":"","institution":"Massachusetts General Hospital, Harvard Medical School","correspondingAuthor":false,"prefix":"","firstName":"Ziya","middleName":"","lastName":"Isiksacan","suffix":""},{"id":407320364,"identity":"3267d00b-bfa1-48f9-ae28-908be66bebbc","order_by":2,"name":"Nishaka William","email":"","orcid":"","institution":"University of Alberta","correspondingAuthor":false,"prefix":"","firstName":"Nishaka","middleName":"","lastName":"William","suffix":""},{"id":407320365,"identity":"bdfa399f-cbe8-493b-9c1d-1719acbaafa0","order_by":3,"name":"Rahime Senturk","email":"","orcid":"","institution":"Massachusetts General Hospital, Harvard Medical School","correspondingAuthor":false,"prefix":"","firstName":"Rahime","middleName":"","lastName":"Senturk","suffix":""},{"id":407320366,"identity":"47d8db6f-2527-4aff-a5e4-b482173a137b","order_by":4,"name":"Luke E. 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Cold storage at +\u0026thinsp;4\u0026deg;C for up to 42 days can reduce transfusion efficacy due to alterations termed the \"storage lesion.\" Strategies to mitigate the storage lesion include alkaline additive solutions and supercooled storage to extend storage by reducing metabolic stresses. However, RBC metabolism during supercooled storage in standard or alkaline additives remains unstudied. This study, thus, investigated the impact of storage additives (alkaline E-Sol5 and standard SAGM) and temperatures (+\u0026thinsp;4\u0026deg;C, -4\u0026deg;C, -8\u0026deg;C) on RBC metabolism during 21- and 42-days storage using high-throughput metabolomics. RBCs stored with E-Sol5 showed increased glycolysis and higher ratios of reduced to oxidized glutathione compared to SAGM. Supercooled storage at -4\u0026deg;C showed markedly lower hemolysis than \u0026minus;\u0026thinsp;8\u0026deg;C, preserved adenylate pools, decreased glucose consumption, and reduced lactate accumulation and pentose phosphate pathway activation. The combination of supercooled storage and E-Sol5 helped to preserve ATP and 2,3-DPG reservoirs, while preventing catabolism and free fatty acid accumulation. While supercooled storage with E-Sol5 offers a promising alternative to standard storage, preserving RBC metabolic and functional quality, further research is necessary to validate and improve on these foundational findings.​​\u003c/p\u003e","manuscriptTitle":"Supercooled storage of red blood cells slows down the metabolic storage lesion","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-02-27 21:22:14","doi":"10.21203/rs.3.rs-5256734/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-05-26T15:36:11+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-23T11:57:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"230675011667261174355752316872448406588","date":"2025-05-07T10:31:04+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-01-26T18:22:16+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"104613647512338944434132116502568809372","date":"2025-01-17T10:05:00+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-11-10T19:55:50+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-11-10T19:50:49+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-10-30T14:19:09+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-10-28T08:33:09+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-10-13T19:46:59+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"220447f4-86df-4a26-a7f0-21d24be689ee","owner":[],"postedDate":"February 27th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":43433206,"name":"Biological sciences/Biotechnology/Metabolomics"},{"id":43433207,"name":"Biological sciences/Biotechnology"},{"id":43433208,"name":"Health sciences/Medical research"},{"id":43433209,"name":"Physical sciences/Engineering"}],"tags":[],"updatedAt":"2025-10-06T16:07:12+00:00","versionOfRecord":{"articleIdentity":"rs-5256734","link":"https://doi.org/10.1038/s41598-025-18028-4","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-10-03 15:57:05","publishedOnDateReadable":"October 3rd, 2025"},"versionCreatedAt":"2025-02-27 21:22:14","video":"","vorDoi":"10.1038/s41598-025-18028-4","vorDoiUrl":"https://doi.org/10.1038/s41598-025-18028-4","workflowStages":[]},"version":"v1","identity":"rs-5256734","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5256734","identity":"rs-5256734","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}