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While several studies have shown an association between L1CAM and glioblastoma (GBM) cell motility, its mechanobiological function in patient\u2010derived primary cultured GBM cells remains insufficiently understood. This study focuses on L1CAM\u2010dependent motility mechanisms in patient\u2010derived GBM cells at the single\u2010cell level. Patient\u2010derived GBM cells were established from GBM tissues using the neurosphere method. L1CAM expression was evaluated by qPCR, western blotting, flow cytometry, and immunocytochemistry. Cell motility was assessed by single\u2010cell tracking. Functional assays included forced expression and antibody\u2010mediated inhibition of L1CAM. Adhesion\u2010clutch behavior and retrograde F\u2010actin flow were analyzed. For in\u00a0vivo assessment, doxycycline\u2010inducible L1CAM\u2010overexpressing GBM cells were transplanted into mouse brains. L1CAM\u2010high GBM cells showed increased motility on laminin compared to L1CAM\u2010low cells. Anti\u2010L1CAM antibody suppressed migration of L1CAM\u2010high GBM cells, whereas ectopic expression enhanced migration in L1CAM\u2010low GBM cells. L1CAM\u2010positive GBM cells preferentially migrated on laminin rather than fibronectin, while L1CAM\u2010low cells showed no such difference, indicating L1CAM\u2010dependent haptotaxis. Forced L1CAM expression reduced retrograde actin flow velocity and promoted adhesion\u2010clutch formation. In\u00a0vivo, GBM cells with L1CAM overexpression displayed greater infiltrative capacity 12\u2009weeks posttransplantation. L1CAM expression on GBM cells' surface regulates cell motility through adhesion\u2010clutch mechanisms and substrate sensing. These mechanobiological features suggest that targeting L1CAM may serve as a promising strategy to suppress GBM cells infiltration and improve GBM treatment outcomes.<\/jats:p>","DOI":"10.1111\/cas.70359","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T23:52:44Z","timestamp":1773013964000},"page":"1380-1396","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["L1CAM Promotes the Infiltrative Properties of Patient\u2010Derived Glioblastoma Cells"],"prefix":"10.1111","volume":"117","author":[{"given":"Asako","family":"Katsuma","sequence":"first","affiliation":[{"name":"Division of Regenerative Medicine, Department of Biomedical Research and Innovation, Institute for Clinical Research NHO Osaka National Hospital  Osaka Japan"},{"name":"Laboratory of Systems Neurobiology and Medicine, Division of Biological Science Nara Institute of Science and Technology  Ikoma Nara 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