Respiring cultureware for high-density, scalable, multipurpose cell-based bioproduction

Respiring cultureware for high-density, scalable, multipurpose cell-based bioproduction | 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 Respiring cultureware for high-density, scalable, multipurpose cell-based bioproduction Colin Cook, Austin Santiago, Nicholas Scianmarello, Seonah Kang, and 14 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8526967/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract In vitro tissue culture remains inefficient due to inferior oxygen transport in polystyrene versus native capillary beds. Taking a bioinspired approach, we engineered respiring cultureware capable of high density, 3D cell culture. Leveraging the oxygen permeability of silicone and finite element modelling, we designed micromolded membranes that provide high oxygen transport (local k L a equivalent > 100/hr). The “high density cell respirator” (HDCR) microarchitecture comprises rows of silicone fins that protrude up from a base membrane. The fins act as artificial capillaries to oxygenate the niche between them, where cells expand. Cellularities of > 1E8 cells/cm 3 are routinely achieved across common cell lines, approaching theoretical limits of 3D confluence. HDCR cultureware is compatible with adherent, suspension, microcarrier, and spheroid cultures, and inherently linearly scalable due to the conserved geometry across 96-well, 24-well, and dish formats. Applications are explored across general cell culture, CAR-T, viral vector, and antibody, demonstrating utility for multipurpose, intensified bioproduction. Biological sciences/Biotechnology Health sciences/Medical research/Translational research Scientific community and society/Scientific community/Lab life Full Text Additional Declarations Yes there is potential Competing Interest. C.C., A.S., N.S., A.C., H.S., G.Z., H.L., N.B., C.T. are or were employees of XDemics Corporation. C.C., A.S., N.S., A.C., H.S., G.Z., N.B., Y.-C.T., Y.F., C.T. hold equity in the company. C.C., A.S., N.S., G.Z., Y.L., S.C., Y.-C.T., Y.F., C.T. are inventors on patent application(s) related to the HDCR technology described in this manuscript. Patent application(s) related to this work have been licensed from City of Hope and California Institute of Technology by XDemics Corporation for commercial development with royalties payable to C.C., Y.L., S.C., Y.-C.T., Y.F.. Cite Share Download PDF Status: Under Review 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8526967","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":577583203,"identity":"43f7cabf-a57f-4102-99e9-2998ba438c5a","order_by":0,"name":"Colin 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C.C., A.S., N.S., A.C., H.S., G.Z., N.B., Y.-C.T., Y.F., C.T. hold equity in the company. C.C., A.S., N.S., G.Z., Y.L., S.C., Y.-C.T., Y.F., C.T. are inventors on patent application(s) related to the HDCR technology described in this manuscript. Patent application(s) related to this work have been licensed from City of Hope and California Institute of Technology by XDemics Corporation for commercial development with royalties payable to C.C., Y.L., S.C., Y.-C.T., Y.F..","formattedTitle":"Respiring cultureware for high-density, scalable, multipurpose cell-based bioproduction","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-8526967/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8526967/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cem\u003eIn vitro\u003c/em\u003e tissue culture remains inefficient due to inferior oxygen transport in polystyrene versus native capillary beds. Taking a bioinspired approach, we engineered respiring cultureware capable of high density, 3D cell culture. Leveraging the oxygen permeability of silicone and finite element modelling, we designed micromolded membranes that provide high oxygen transport (local \u003cem\u003ek\u003c/em\u003e\u003csub\u003eL\u003c/sub\u003ea equivalent\u0026thinsp;\u0026gt;\u0026thinsp;100/hr). The \u0026ldquo;high density cell respirator\u0026rdquo; (HDCR) microarchitecture comprises rows of silicone fins that protrude up from a base membrane. The fins act as artificial capillaries to oxygenate the niche between them, where cells expand. Cellularities of \u0026gt;\u0026thinsp;1E8 cells/cm\u003csup\u003e3\u003c/sup\u003e are routinely achieved across common cell lines, approaching theoretical limits of 3D confluence. HDCR cultureware is compatible with adherent, suspension, microcarrier, and spheroid cultures, and inherently linearly scalable due to the conserved geometry across 96-well, 24-well, and dish formats. Applications are explored across general cell culture, CAR-T, viral vector, and antibody, demonstrating utility for multipurpose, intensified bioproduction.\u003c/p\u003e","manuscriptTitle":"Respiring cultureware for high-density, scalable, multipurpose cell-based bioproduction","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-11 11:13:35","doi":"10.21203/rs.3.rs-8526967/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"communications-biology","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"commsbio","sideBox":"Learn more about [Communications Biology](http://www.nature.com/commsbio/)","snPcode":"","submissionUrl":"","title":"Communications Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Communications Series","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"7fcd1cc4-7ab6-431a-b5a1-78d9de8a5c3f","owner":[],"postedDate":"February 11th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":61449391,"name":"Biological sciences/Biotechnology"},{"id":61449392,"name":"Health sciences/Medical research/Translational research"},{"id":61449393,"name":"Scientific community and society/Scientific community/Lab life"}],"tags":[],"updatedAt":"2026-03-26T08:00:13+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-11 11:13:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8526967","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8526967","identity":"rs-8526967","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}