Safety of Hypoxia-Primed Mesenchymal Stem Cells Derived from Umbilical Cord and Adipose Tissuesin Animals

Safety of Hypoxia-Primed Mesenchymal Stem Cells Derived from Umbilical Cord and Adipose Tissuesin Animals | 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 Research Article Safety of Hypoxia-Primed Mesenchymal Stem Cells Derived from Umbilical Cord and Adipose Tissuesin Animals Nhung Thi-Hong Dinh, Quyen Thi Nguyen, Ngo Thu Hang, Hong-Nhung Dao, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5182044/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Mesenchymal stem cells (MSCs) have been applied to treat various diseases. Research has shown that hypoxic conditions (2-9% oxygen) that closely reflect the physiological milieu of numerous human tissues, can enhance the biological activities of MSCs in vitro. While many studies highlight the advantages of hypoxic MSCs in vitro , less is known about their effects in vivo . This study assessed the safety of hypoxia-primed MSCs derived from adipose tissues and umbilical cords (AD-MSCs and UC-MSCs, respectively) in healthy animals. Methods: MSCs were isolated from human adipose tissues and umbilical cords, designated as AD-MSCs and UC-MSCs, respectively. These cells were cultured under hypoxic conditions with 5% oxygen to induce priming. For safety evaluation, the primed cells were administered to healthy animal models, including rabbits, Swiss mice, and rats. The safety assessments included analyses of vascular and muscular stimulation, systemic hypersensitivity, acute toxicity, and subchronic toxicity. Results: Our data indicated that UC-MSCs induced vascular stimulation but not muscular stimulation, whereas AD-MSCs caused neither. Additionally, neither AD-MSCs nor UC-MSCs affected hematopoietic parameters (white blood cell, red blood cell, platelet, or hemoglobin levels), altered the levels of proinflammatory cytokines (IL-6, IL-1β, IFN-γ, and TNF-α), or induced allergenic responses in rabbits. Furthermore, intravenous injection of either UC-MSCs or AD-MSCs at a dose of 50 × 10 6 cells/kg did not cause acute toxicity in mice. However, injections of higher doses of these cells led to intravenous thrombosis and embolism in various organs of experimental animals, ultimately resulting in animal death. Finally, according to the subchronic toxicity assay, the administration of MSCs generally did not impact liver, kidney, or spleen function in rats. Conclusions: Hypoxic MSCs are safe for therapeutic use with consideration of thrombogenic risk and could be alternatives to normoxic MSCs for disease treatment. Mesenchymal stem cells (MSCs) hypoxia-primed culture safety evaluation vascular and muscle stimulation systemic hypersensitivity acute toxicity subchronic toxicity Full Text Additional Declarations The authors declare no competing interests. Supplementary Files Additionalfile1.docx Cite Share Download PDF Status: Posted Version 1 posted 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. We do this by developing innovative software and high quality services for the global research community. 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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-5182044","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":360733597,"identity":"1975e2b9-ed4b-440f-b3aa-b662cdf623c6","order_by":0,"name":"Nhung Thi-Hong Dinh","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Nhung","middleName":"Thi-Hong","lastName":"Dinh","suffix":""},{"id":360734627,"identity":"ba91567e-af2f-48c4-975f-1cf2a75cd371","order_by":1,"name":"Quyen Thi 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Animals\u003c/strong\u003e\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Vinmec Research Institute of Stem Cell and Gene Technology","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Mesenchymal stem cells (MSCs), hypoxia-primed culture, safety evaluation, vascular and muscle stimulation, systemic hypersensitivity, acute toxicity, subchronic toxicity","lastPublishedDoi":"10.21203/rs.3.rs-5182044/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5182044/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eMesenchymal stem cells (MSCs) have been applied to treat various diseases. Research has shown that hypoxic conditions (2-9% oxygen) that closely reflect the physiological milieu of numerous human tissues, can enhance the biological activities of MSCs \u003cem\u003ein vitro. \u003c/em\u003eWhile many studies highlight the advantages of hypoxic MSCs \u003cem\u003ein vitro\u003c/em\u003e, less is known about their effects \u003cem\u003ein vivo\u003c/em\u003e. This study assessed the safety of hypoxia-primed MSCs derived from adipose tissues and umbilical cords (AD-MSCs and UC-MSCs, respectively) in healthy animals.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eMSCs were isolated from human adipose tissues and umbilical cords, designated as AD-MSCs and UC-MSCs, respectively. These cells were cultured under hypoxic conditions with 5% oxygen to induce priming. For safety evaluation, the primed cells were administered to healthy animal models, including rabbits, Swiss mice, and rats. The safety assessments included analyses of vascular and muscular stimulation, systemic hypersensitivity, acute toxicity, and subchronic toxicity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eOur data indicated that UC-MSCs induced vascular stimulation but not muscular stimulation, whereas AD-MSCs caused neither. Additionally, neither AD-MSCs nor UC-MSCs affected hematopoietic parameters (white blood cell, red blood cell, platelet, or hemoglobin levels), altered the levels of proinflammatory cytokines (IL-6, IL-1β, IFN-γ, and TNF-α), or induced allergenic responses in rabbits. Furthermore, intravenous injection of either UC-MSCs or AD-MSCs at a dose of 50 × 10\u003csup\u003e6\u003c/sup\u003e cells/kg did not cause acute toxicity in mice. However, injections of higher doses of these cells led to intravenous thrombosis and embolism in various organs of experimental animals, ultimately resulting in animal death. Finally, according to the subchronic toxicity assay, the administration of MSCs generally did not impact liver, kidney, or spleen function in rats.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions: \u003c/strong\u003eHypoxic MSCs are safe for therapeutic use with consideration of thrombogenic risk and could be alternatives to normoxic MSCs for disease treatment.\u003c/p\u003e","manuscriptTitle":"Safety of Hypoxia-Primed Mesenchymal Stem Cells Derived from Umbilical Cord and Adipose Tissuesin Animals","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-03 03:20:46","doi":"10.21203/rs.3.rs-5182044/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4a8a239a-7449-4795-b4aa-0e3dbafe1fea","owner":[],"postedDate":"October 3rd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-10-03T03:20:46+00:00","versionOfRecord":[],"versionCreatedAt":"2024-10-03 03:20:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5182044","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5182044","identity":"rs-5182044","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}