{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:42:07Z","timestamp":1760060527102,"version":"build-2065373602"},"reference-count":44,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,1]],"date-time":"2025-09-01T00:00:00Z","timestamp":1756684800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Grants-in-aid for Japan Society for the Promotion of Science","award":["20K07773","24K10032"],"award-info":[{"award-number":["20K07773","24K10032"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Neurology International"],"abstract":"<jats:p>Background\/Objectives: Co-culture models of neurons and Schwann cells have been used to explore the mechanisms of myelination during development, axonal regeneration after injury, and the pathogenesis of various demyelinating neuropathies. A spontaneously immortalized Fischer rat Schwann cell line 1 (IFRS1), established from the primary culture of adult Fischer344 rat peripheral nerves, can myelinate neurites in co-cultures with primary cultured dorsal root ganglion neurons and neuronal cell lines, such as nerve growth factor (NGF)-primed PC12 cells and NSC-34 motor neuron-like cells. In this study, we aimed to establish a stable co-culture system using IFRS1 cells and ND7\/23 sensory neuron-like cells. Methods: ND7\/23 cells were seeded at a low density (2 \u00d7 103\/cm2) and maintained for 7 days in serum-containing medium supplemented with NGF (10 ng\/mL) and the Rho kinase inhibitor Y27632 (5 \u03bcM) to promote neurite elongation. The cells were then treated with the anti-mitotic agent mitomycin C (1 \u03bcg\/mL) for 12\u201316 h to suppress proliferative activity. Following this, the cells were co-cultured with IFRS1 cells (2 \u00d7 104\/cm2) and maintained at 37 \u00b0C in serum-containing medium supplemented with ascorbic acid (50 \u03bcg\/mL), NGF (10 ng\/mL), and ciliary neurotrophic factor (10 ng\/mL). Results: Double-immunofluorescence staining performed on day 21 of the co-culture revealed myelin protein 22- or myelin basic protein-immunoreactive IFRS1 cells surrounding \u03b2III tubulin-immunoreactive neurites emerging from ND7\/23 cells. Myelin formation was further confirmed via Sudan Black B staining and electron microscopy. Conclusions: This co-culture system may provide a valuable tool for studying the processes of myelination in the peripheral nervous system, as well as the pathogenesis of various sensory neuropathies and potential novel therapeutic approaches for these conditions.<\/jats:p>","DOI":"10.3390\/neurolint17090138","type":"journal-article","created":{"date-parts":[[2025,9,1]],"date-time":"2025-09-01T13:27:44Z","timestamp":1756733264000},"page":"138","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Effects of Co-Culturing ND7\/23 Sensory Neuron-like Cells and IFRS1 Schwann Cells on Myelination: A Single-Arm Nonrandomized Study"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8496-5181","authenticated-orcid":false,"given":"Shizuka","family":"Takaku","sequence":"first","affiliation":[{"name":"Diabetic Neuropathy Project, Department of Diseases and Infection, Tokyo Metropolitan Institute of Medical Science, Setagaya-ku, Tokyo 156-8506, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9750-9596","authenticated-orcid":false,"given":"Kazunori","family":"Sango","sequence":"additional","affiliation":[{"name":"Diabetic Neuropathy Project, Department of Diseases and Infection, Tokyo Metropolitan Institute of Medical Science, Setagaya-ku, Tokyo 156-8506, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1007\/s00441-025-03957-3","article-title":"The role and mechanism of Schwann cells in the repair of peripheral nerve injury","volume":"400","author":"Xu","year":"2025","journal-title":"Cell Tissue Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1002\/glia.23509","article-title":"The conceptual introduction of the \u201cdemyelinating Schwann cell\u201d in peripheral demyelinating neuropathies","volume":"67","author":"Park","year":"2019","journal-title":"Glia"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Schenone, A., Massucco, S., Schenone, C., Venturi, C.B., Nozza, P., Prada, V., Pomili, T., Di Patrizi, I., Capodivento, G., and Nobbio, L. 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