Abstract
Dramatic progress has been made over recent years toward the elucidation of the mechanisms regulating lineage determination and cell survival in the developing peripheral nervous system. However, our understanding of Schwann cell development is limited. This is partly due to the difficulties in culturing primary Schwann cell precursor cells, the earliest developmental stage of the Schwann cell lineage defined to date. Both the inability to maintain cultured Schwann cell precursor cells in an undifferentiated state and the technical difficulties involved in their isolation have hampered progress. We have conditionally immortalized rat Schwann cell precursor cells using a retrovirally encoded EGFR/neu fusion protein to circumvent these problems and to generate a source of homogeneous cells. The resulting SpL201 cell line expresses p75 and nestin, two proteins expressed by neural crest-derived cells, as well as peripheral myelin protein 22, protein zero, and Oct-6 as markers of the Schwann cell lineage. When cultured in EGF-containing medium, the SpL201 cells proliferate and maintain an undifferentiated, Schwann cell precursor cell-like state. The cell line is dependent on EGF for survival but can differentiate into early Schwann cell-like cells in response to exogenous factors. Like primary rat Schwann cells, SpL201 cells upregulate Oct-6 and myelin gene expression in response to forskolin treatment. Furthermore, the SpL201 cell line can form myelin in the presence of axons in vitro and is capable of extensively remyelinating a CNS white matter lesion in vivo. Thus, this cell line provides a valuable and unique tool to study the Schwann cell lineage, including differentiation from the Schwann cell precursor cell stage through to myelination.
Copyright 2001 Wiley-Liss, Inc.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigens, Differentiation / metabolism
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Blood Proteins / pharmacology
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Calcium-Binding Proteins / metabolism
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Cell Culture Techniques / methods
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Cell Differentiation / drug effects
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Cell Differentiation / physiology
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Cell Line, Transformed / metabolism*
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Cell Line, Transformed / ultrastructure
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Cell Lineage / physiology
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Cell Survival / drug effects
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Cell Survival / physiology
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Colforsin / pharmacology
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Culture Media, Conditioned / pharmacology
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Epidermal Growth Factor / pharmacology
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Fibroblasts / drug effects
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Fibroblasts / metabolism
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Immunohistochemistry
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Intermediate Filament Proteins / metabolism
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Microscopy, Electron
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Mitogens / pharmacology
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Myelin Proteins / drug effects
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Myelin Proteins / metabolism
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Myelin Sheath / metabolism*
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Nerve Growth Factors / metabolism
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Nerve Regeneration / physiology
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Nerve Tissue Proteins*
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Nestin
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Neural Crest / cytology
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Neural Crest / embryology
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Neural Crest / metabolism
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Neurites / ultrastructure
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Octamer Transcription Factor-6
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Rats
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Rats, Sprague-Dawley
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Receptor, Nerve Growth Factor
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Receptors, Nerve Growth Factor / metabolism
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S100 Calcium Binding Protein beta Subunit
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S100 Proteins*
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Schwann Cells / metabolism*
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Schwann Cells / ultrastructure
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Stem Cells / metabolism*
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Stem Cells / ultrastructure
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Transcription Factors / drug effects
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Transcription Factors / metabolism
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Up-Regulation / drug effects
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Up-Regulation / physiology
Substances
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Antigens, Differentiation
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Blood Proteins
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Calcium-Binding Proteins
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Culture Media, Conditioned
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Intermediate Filament Proteins
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Mitogens
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Myelin Proteins
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Nerve Growth Factors
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Nerve Tissue Proteins
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Nes protein, mouse
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Nes protein, rat
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Nestin
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Pmp22 protein, mouse
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Pmp22 protein, rat
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Pou3f1 protein, rat
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Receptor, Nerve Growth Factor
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Receptors, Nerve Growth Factor
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S100 Calcium Binding Protein beta Subunit
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S100 Proteins
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Transcription Factors
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oligodendrocyte O antigen, rat
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Octamer Transcription Factor-6
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Colforsin
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Epidermal Growth Factor