Document Detail


A novel method for isolating Schwann cells using the extracellular domain of Necl1.
MedLine Citation:
PMID:  19125407     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Myelinating cocultures of Schwann cells and dorsal root ganglion neurons are a powerful experimental system for probing the molecular mechanisms of axon-Schwann cell interaction. The isolation of a pure population of myelination-competent Schwann cells is a prerequisite for this experimental system. We describe here a protocol for a FACS-based isolation of Schwann cells utilizing a specific affinity reagent (Necl1-Fc) and the use of these isolated cells in myelinating cocultures. An advantage of the myelinating coculture system is that Schwann cells and the neurons can be genetically manipulated before they are cocultured. We further show that our method allows the isolation of virally transduced Schwann cells in a single purification step. This protocol for the FACS-based isolation of myelination-competent Schwann cells by Necl1-Fc and the use of these cells in myelinating cocultures should significantly facilitate future studies aimed at delineation of the molecular mechanisms of axon-Schwann cell interactions and myelination.
Authors:
Ivo Spiegel; Elior Peles
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Journal of neuroscience research     Volume:  87     ISSN:  1097-4547     ISO Abbreviation:  J. Neurosci. Res.     Publication Date:  2009 Nov 
Date Detail:
Created Date:  2009-10-12     Completed Date:  2010-02-01     Revised Date:  2010-09-23    
Medline Journal Info:
Nlm Unique ID:  7600111     Medline TA:  J Neurosci Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3288-96     Citation Subset:  IM    
Affiliation:
Department of Molecular Cell Biology, The Weizmann Institute of Science, Rehovot, Israel.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Adhesion Molecules, Neuronal / chemistry*,  metabolism
Cell Communication / physiology
Cell Culture Techniques / methods
Cell Line
Cell Membrane / chemistry,  metabolism
Cells, Cultured
Coculture Techniques
Culture Media, Conditioned / chemistry,  metabolism
Flow Cytometry / methods*
Ganglia, Spinal / cytology,  metabolism
Genetic Vectors / pharmacology
Humans
Immunoglobulin G / chemistry,  metabolism
Neurons / cytology,  metabolism
Neurosciences / methods*
Protein Binding / physiology
Protein Structure, Tertiary / physiology
Rats
Recombinant Fusion Proteins / chemistry*,  metabolism
Schwann Cells / cytology,  metabolism*
Transduction, Genetic / methods
Grant Support
ID/Acronym/Agency:
NS50220/NS/NINDS NIH HHS; R01 NS050220-04/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Cell Adhesion Molecules, Neuronal; 0/Culture Media, Conditioned; 0/Immunoglobulin G; 0/Recombinant Fusion Proteins; 0/nectin-like molecule 1, rat
Comments/Corrections

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