Generation and characterization of human induced pluripotent stem cell lines from patients with autism spectrum disorder and SCN2A variants

Generation and characterization of human induced pluripotent stem cell lines from patients with autism spectrum disorder and SCN2A variants | 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 Generation and characterization of human induced pluripotent stem cell lines from patients with autism spectrum disorder and SCN2A variants John Lenon de Souza Santos, Bruno Diaz Paredes, Corynne Stephanie Ahouefa Adanho, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4676288/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 20 Mar, 2025 Read the published version in Human Cell → Version 1 posted 5 You are reading this latest preprint version Abstract Autism spectrum disorders (ASD) comprise a group of complex neurodevelopmental disorders that affect communication and social interactions. Over a thousand genes have been associated with ASD, with SCN2A standing out due to its critical role in neuronal function and development. Induced pluripotent stem cells (iPSCs) derived from individuals with ASD have become invaluable in vitro models for investigating the cellular and molecular mechanisms underlying the disorder. In this study, we generated and characterized four iPSC clones from peripheral blood mononuclear cells (PBMCs) of two ASD patients carrying loss-of-function variants in the SCN2A gene. These iPSC lines underwent comprehensive characterization through multiple assays. Reverse transcription polymerase chain reaction (RT-PCR), flow cytometry, and immunofluorescence analyses confirmed the presence of pluripotency markers. An embryoid body formation assay demonstrated their potential to differentiate into the three germ layers. Sequencing analysis confirmed the SCN2A variants, while short tandem repeat (STR) analysis authenticated the cell lines, and karyotype analysis ensured chromosomal integrity. The iPSCs exhibited typical morphological characteristics, including large nuclei with prominent nucleoli, a high nucleus-to-cytoplasm ratio, densely packed cells, and well-defined borders. These cells maintained pluripotency markers, demonstrated the ability to differentiate into the three germ layers, and showed a normal karyotype. Furthermore, we successfully generated cerebral organoids from these cells. Our study establishes a robust platform for further exploration of the pathophysiological mechanisms of ASD, particularly those involving SCN2A. iPSCs SCN2A variants Autism spectrum disorder brain organoids Full Text Supplementary Files Supplementary.docx Cite Share Download PDF Status: Published Journal Publication published 20 Mar, 2025 Read the published version in Human Cell → Version 1 posted Editorial decision: Major Revisions Needed 24 Nov, 2024 Reviewers agreed at journal 22 Jul, 2024 Reviewers invited by journal 06 Jul, 2024 Editor assigned by journal 04 Jul, 2024 First submitted to journal 02 Jul, 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. 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-4676288","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":323466300,"identity":"1e077d37-e2f2-41f7-b6cf-6d2c2aa00798","order_by":0,"name":"John Lenon de Souza Santos","email":"","orcid":"","institution":"FIOCRUZ Bahia: Instituto Goncalo Moniz","correspondingAuthor":false,"prefix":"","firstName":"John","middleName":"Lenon de Souza","lastName":"Santos","suffix":""},{"id":323466301,"identity":"a7681abc-087b-4c5e-81de-bacdd5a0ca04","order_by":1,"name":"Bruno Diaz Paredes","email":"","orcid":"","institution":"IDOR: Instituto D'Or 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