An exploration of vascular cell development in mouse embryoid bodies under hypoxia through single cell RNA-seq

An exploration of vascular cell development in mouse embryoid bodies under hypoxia through single cell RNA-seq | 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 An exploration of vascular cell development in mouse embryoid bodies under hypoxia through single cell RNA-seq Bárbara Acosta-Iborra, Laura Puente-Santamaría, Yosra Berrouayel, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3954034/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 : Differentiation of mouse stem cells (mESC) to embryonic bodies (EBs) is a widely used approach to model early stages of development. In this study, we employed single-cell transcriptomics to examine the effect of hypoxia on the differentiation of different cell populations in EBs. Results : We differentiated EBs for 8 or 10 days, with two time frames of hypoxic or normoxic exposure: a short window of 16h for the 8-day samples, and a 48h window for the 10-day samples, totaling four experimental conditions. The EBs were composed mainly of broad mesodermal progenitors, a limited number of endodermal precursors, and smaller groups of cells that were more advanced in their differentiation pathways. These developed groups included hemoendothelial progenitors, endothelium, cardiomyocytes, and monocyte and erythrocyte precursors. Both O2 and time of differentiation significantly influenced the distribution of cell types across conditions. Our findings revealed that hypoxia induces a pervasive cell cycle arrest signature across all identified cell populations within the EB. Despite the cell cycle arrest, vascular smooth muscle (vSMCs), endothelial, and cardiac cell populations exhibit expansion under hypoxic conditions, while normoxia favours an increase in the monocyte population. Differential proliferation of hemoendothelial progenitors does not appear to be the primary driver of endothelial cells expansion in hypoxia, as their distribution remained similar across treatments and depleted over time. We observed delays in the terminal differentiation pathway of vSMCs precursors in hypoxic conditions, which may contribute to their expansion and migration by switching to a synthetic phenotype. Additionally, oxygen concentration influenced the developmental path of hemangioblasts towards either an endothelial, or myeloid lineage. In the EBs, this was coupled with a shift in presomitic mesoderm differentiation towards endotome or musculoskeletal precursors, which may provide an additional source of progenitors with endothelial potential during oxygen deprivation. Conclusion : Our study provides novel insights into the cellular responses to hypoxia in the context of early embryonic development, thus unveiling potential mechanisms underlying blood vessel growth. Hypoxia Embryoid Body scRNA-seq Full Text Additional Declarations No competing interests reported. Supplementary Files Additionalfile1.xlsx Additionalfile2.xlsx Additionalfile3.xlsx Additionalfile4.xlsx Additionalfile5.xlsx Additionalfile6.xlsx 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. 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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-3954034","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":274791617,"identity":"ed4887d1-983c-4d2e-895e-7f97bd975cf7","order_by":0,"name":"Bárbara Acosta-Iborra","email":"","orcid":"","institution":"Universidad Autónoma de Madrid (UAM)","correspondingAuthor":false,"prefix":"","firstName":"Bárbara","middleName":"","lastName":"Acosta-Iborra","suffix":""},{"id":274791618,"identity":"222b9fc5-f8f5-4143-a875-a5fdfb0348e0","order_by":1,"name":"Laura 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