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Myelinated, synapsing cultures of murine spinal cord - validation as an in vitro model of the central nervous system

Thomson, C.E. and McCulloch, M. and Sorensen, Annette and Barnett, S.C. and Seed, B.V. and Griffiths, I.R. and McLaughlin, M. (2008) Myelinated, synapsing cultures of murine spinal cord - validation as an in vitro model of the central nervous system. European Journal of Neuroscience, 28 (8). pp. 1518-1535. ISSN 0953-816X

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Abstract

Research in central nervous system (CNS) biology and pathology requires in vitro models, which, to recapitulate the CNS in vivo, must have extensive myelin and synapse formation under serum-free (defined) conditions. However, finding such a model has proven difficult. The technique described here produces dense cultures of myelinated axons, with abundant synapses and nodes of Ranvier, that are suitable for both morphological and biochemical analysis. Cellular and molecular events were easily visualised using conventional microscopy. Ultrastructurally, myelin sheaths were of the appropriate thickness relative to axonal diameter (G-ratio). Production of myelinated axons in these cultures was consistent and repeatable, as shown by statistical analysis of multiple experimental repeats. Myelinated axons were so abundant that from one litter of embryonic mice, myelin was produced in amounts sufficient for bulk biochemical analysis. This culture method was assessed for its ability to generate an in vitro model of the CNS that could be used for both neurobiological and neuropathological research. Myelin protein kinetics were investigated using a myelin fraction isolated from the cultures. This fraction was found to be superior, quantitatively and qualitatively, to the fraction recovered from standard cultures of dissociated oligodendrocytes, or from brain slices. The model was also used to investigate the roles of specific molecules in the pathogenesis of inflammatory CNS diseases. Using the defined conditions offered by this culture system, dose-specific, inhibitory effects of inflammatory cytokines on myelin formation were demonstrated, unequivocally. The method is technically quick, easy and reliable, and should have wide application to CNS research.

Item type: Article
ID code: 26028
Keywords: cytokines, MAG, multiple sclerosis, PLP-DM20, western blotting, spinal cord, Other systems of medicine, Neuroscience(all)
Subjects: Medicine > Other systems of medicine
Department: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
Unknown Department
Related URLs:
    Depositing user: Strathprints Administrator
    Date Deposited: 24 Aug 2010 12:27
    Last modified: 05 Sep 2014 05:10
    URI: http://strathprints.strath.ac.uk/id/eprint/26028

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