Assessing the distribution of cancer stem cells in tumorspheres

Assessing the distribution of cancer stem cells in tumorspheres | 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 Assessing the distribution of cancer stem cells in tumorspheres Jerónimo Fotinós, María Paula Marks, Lucas Barberis, Luciano Vellón This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4004487/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 8 You are reading this latest preprint version Abstract In previous theoretical research, we inferred that cancer stem cells (CSCs), the cells that presumably drive tumor growth and resistance to conventional cancer treatments, are not uniformly distributed in the bulk of a tumorsphere. To confirm this theoretical prediction, we cultivated tumorspheres enriched in CSCs, and performed immunofluorecent detection of the stemness marker SOX2 using a confocal microscope. In this article, we present a method developed to process the images that reconstruct the amount and location of the CSCs in the tumorspheres. Its advantage is the use of a statistical criterion to classify the cells in stem and differentiated instead of setting an arbitrary threshold. From the analysis of the results of the methods using graph theory and computational modeling, we concluded that the distribution of Cancer Stem Cells in an experimental tumorsphere is non-homogeneous. This method is independent of the tumorsphere assay being useful for analyzing images in which several different kinds of cells are stained with different markers. Biological sciences/Biophysics Biological sciences/Computational biology and bioinformatics Physical sciences/Mathematics and computing Physical sciences/Mathematics and computing/Scientific data Biological sciences/Cancer Biological sciences/Cancer/Cancer models Biological sciences/Cancer/Cancer stem cells Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 08 Apr, 2024 Reviews received at journal 02 Apr, 2024 Reviewers agreed at journal 20 Mar, 2024 Reviewers invited by journal 20 Mar, 2024 Editor assigned by journal 14 Mar, 2024 Editor invited by journal 11 Mar, 2024 Submission checks completed at journal 11 Mar, 2024 First submitted to journal 01 Mar, 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. 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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-4004487","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":278570187,"identity":"a6f7b29a-1428-4c8a-9769-eb5ba5faa488","order_by":0,"name":"Jerónimo Fotinós","email":"","orcid":"","institution":"Instituto de Física Enrique Gaviola","correspondingAuthor":false,"prefix":"","firstName":"Jerónimo","middleName":"","lastName":"Fotinós","suffix":""},{"id":278570188,"identity":"ab833f42-68b9-4303-843e-67c0ffb895e3","order_by":1,"name":"María Paula Marks","email":"","orcid":"","institution":"Instituto de Biologia y Medicina Experimental","correspondingAuthor":false,"prefix":"","firstName":"María","middleName":"Paula","lastName":"Marks","suffix":""},{"id":278570189,"identity":"ddb61876-9efa-4608-b4a0-149f935e4428","order_by":2,"name":"Lucas Barberis","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3klEQVRIiWNgGAWjYBACPgkGBmYgzcPA3gAWYGwgpIUNroXnAIlaGBgkEojVIt387HNBxT0Z+ZlvzB78YLCR3XCA+dkHvFpkjhnPnnGmmMfgdo65YQ9DmvGGA2zGM/A7LMGYmbctgcdAOsdMmoHhcOKGAwzGBPyS/pmZ918Cj/zMMyAt/4Fa2D8T0JIDtKUhgYfhBg9IywGgFh4CtsicKWaecQzosDNpZZI9BsnGMw/zFOPVwi/dvpm5oCbBXr798DaJHxV2sn3H2zfj1YIGDBhg0TQKRsEoGAWjgBIAAB9DPjbp4v5QAAAAAElFTkSuQmCC","orcid":"","institution":"Instituto de Física Enrique Gaviola","correspondingAuthor":true,"prefix":"","firstName":"Lucas","middleName":"","lastName":"Barberis","suffix":""},{"id":278570190,"identity":"61f40fc1-5f6a-4e69-8dbd-591f7cbd15ce","order_by":3,"name":"Luciano Vellón","email":"","orcid":"","institution":"Instituto de Biologia y Medicina Experimental","correspondingAuthor":false,"prefix":"","firstName":"Luciano","middleName":"","lastName":"Vellón","suffix":""}],"badges":[],"createdAt":"2024-03-01 20:48:44","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4004487/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4004487/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":52574174,"identity":"cad3789a-1fa9-4973-9171-4adff6921735","added_by":"auto","created_at":"2024-03-13 06:30:25","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4188626,"visible":true,"origin":"","legend":"","description":"","filename":"submission.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4004487/v1_covered_974b39b0-8cfd-4e4f-9b3d-7eae85139598.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Assessing the distribution of cancer stem cells in tumorspheres","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":"","lastPublishedDoi":"10.21203/rs.3.rs-4004487/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4004487/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"In previous theoretical research, we inferred that cancer stem cells (CSCs), the cells that presumably drive tumor growth and resistance to conventional cancer treatments, are not uniformly distributed in the bulk of a tumorsphere. 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