Mechano-Dynamic Brain-on-Chip for Studying Human Stem Cell-Derived Neuronal Networks

Mechano-Dynamic Brain-on-Chip for Studying Human Stem Cell-Derived Neuronal Networks | 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 Mechano-Dynamic Brain-on-Chip for Studying Human Stem Cell-Derived Neuronal Networks Regina Luttge, Gulden Akcay This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6121254/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 13 Aug, 2025 Read the published version in Scientific Reports → Version 1 posted 11 You are reading this latest preprint version Abstract Brain tissue orchestrates neuronal function through biochemical and mechanical cues. Utilizing in vitro modeling, often the dynamics of mechanical aspects in neuronal cell cultures is neglected. However, the growing recognition of the importance of mechanical cues in neural development and healthy brain function necessitates a shift in how we study cultured neurons. Microfluidic platforms, like a Brain-on-Chip (BoC), can take active mechanical stimuli into account. In our BoC design a set of microchannels manufactured in a glass substrate by FEMTOprint technology is assembled with a spin-coated polydimethylsiloxane (PDMS) membrane and a PDMS culture chamber, which was fabricated from a stereolithographically made mold by replication. The membrane can locally deform across the culture chamber by air pressure. This paper describes the design, fabrication and test of such a novel BoC, offering an experimental setting in which we demonstrated mechano-dynamic elevated Calcium signaling in cultured human induced neural stem cell-derived neuronal networks. Physical sciences/Engineering/Biomedical engineering Biological sciences/Neuroscience Brain-on-Chip (BoC) Mechanical stimulation hiPSCs-derived neurons Neuronal activity Calcium live imaging Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryMaterial.pdf Cite Share Download PDF Status: Published Journal Publication published 13 Aug, 2025 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 12 May, 2025 Reviews received at journal 05 May, 2025 Reviews received at journal 30 Apr, 2025 Reviewers agreed at journal 17 Apr, 2025 Reviewers agreed at journal 16 Apr, 2025 Reviewers agreed at journal 16 Apr, 2025 Reviewers invited by journal 13 Apr, 2025 Editor assigned by journal 07 Apr, 2025 Editor invited by journal 11 Mar, 2025 Submission checks completed at journal 07 Mar, 2025 First submitted to journal 07 Mar, 2025 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-6121254","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":430971732,"identity":"4dffc3c8-3937-4c13-a345-04c8f654397c","order_by":0,"name":"Regina Luttge","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAr0lEQVRIiWNgGAWjYFAC5gYwxQ/hWRCjhbGB4UACA4NkG5gnQYIWg2PEajFvYGyT/vijVs74fo/hA8YdRGiROcDYJnEg4bix2TEeYwPGM0RokWAAazmWuO0Yj5kEkE28lvrNbTzmP0jRUpNgwMZjxkCcFmbGZoszaQcMZxxLK5ZIJEoLe/PBGxU2dfL8zYc3fvjYZkNYCwMzmDwM4SQQoQEG6khQOwpGwSgYBSMOAABvijKhTxARKQAAAABJRU5ErkJggg==","orcid":"","institution":"Eindhoven University of Technology","correspondingAuthor":true,"prefix":"","firstName":"Regina","middleName":"","lastName":"Luttge","suffix":""},{"id":430971734,"identity":"e8273be9-5879-48c0-bcd4-32aad1d8a5f4","order_by":1,"name":"Gulden Akcay","email":"","orcid":"","institution":"Eindhoven University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Gulden","middleName":"","lastName":"Akcay","suffix":""}],"badges":[],"createdAt":"2025-02-27 13:08:32","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6121254/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6121254/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-025-14187-6","type":"published","date":"2025-08-13T15:57:09+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":89310533,"identity":"9fa6e9c4-f85a-4fdb-81a2-d9de4dc763f8","added_by":"auto","created_at":"2025-08-18 16:07:14","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":76970879,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscriptMinorRevisions.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6121254/v1_covered_5dc0cfe6-d7ad-4287-89e5-ab63b7812e5f.pdf"},{"id":78938524,"identity":"b247b684-4923-45b5-a7d6-f05669bea13b","added_by":"auto","created_at":"2025-03-21 06:02:54","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":8560598,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterial.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6121254/v1/cf23171274c89f7b319a80cd.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Mechano-Dynamic Brain-on-Chip for Studying Human Stem Cell-Derived Neuronal Networks","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"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},"keywords":"Brain-on-Chip (BoC), Mechanical stimulation, hiPSCs-derived neurons, Neuronal activity, Calcium live imaging","lastPublishedDoi":"10.21203/rs.3.rs-6121254/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6121254/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Brain tissue orchestrates neuronal function through biochemical and mechanical cues. 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