| Myelination in coculture of established neuronal and Schwann cell lines. | |
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MedLine Citation:
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PMID: 22366958 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Establishing stable coculture systems with neuronal and Schwann cell lines has been considered difficult, presumably because of their high proliferative activity and phenotypic differences from primary cultured cells. The present study is aimed at developing methods for myelin formation under coculture of the neural crest-derived pheochromocytoma cell line PC12 and the immortalized adult rat Schwann cell line IFRS1. Prior to coculture, PC12 cells were seeded at low density (3 × 10(2)/cm(2)) and maintained in serum-free medium with N2 supplement, ascorbic acid (50 μg/ml), and nerve growth factor (NGF) (50 ng/ml) for a week. Exposure to such a NGF-rich environment with minimum nutrients accelerated differentiation and neurite extension, but not proliferation, of PC12 cells. When IFRS1 cells were added to NGF-primed PC12 cells, the cell density ratio of PC12 cells to IFRS1 cells was adjusted from 1:50 to 1:100. The cocultured cells were then maintained in serum-free medium with B27 supplement, ascorbic acid (50 μg/ml), NGF (10 ng/ml), and recombinant soluble neuregulin-1 type III (25 ng/ml). Myelin formation was illustrated by light and electron microscopy performed at day 28 of coculture. The stable PC12-IFRS1 coculture system is free of technical and ethical problems arising from the primary culture and can be a valuable tool to study peripheral nerve degeneration and regeneration. |
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Authors:
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Kazunori Sango; Emiko Kawakami; Hiroko Yanagisawa; Shizuka Takaku; Masami Tsukamoto; Kazunori Utsunomiya; Kazuhiko Watabe |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-2-25 |
Journal Detail:
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Title: Histochemistry and cell biology Volume: - ISSN: 1432-119X ISO Abbreviation: - Publication Date: 2012 Feb |
Date Detail:
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Created Date: 2012-2-27 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9506663 Medline TA: Histochem Cell Biol Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
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Department of Sensory and Motor Systems, ALS/Neuropathy Project (Laboratory of Peripheral Nerve Pathophysiology), Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan, sango-kz@igakuken.or.jp. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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