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

Thomson, C.E., McCulloch, M., Sorenson, A., Barnett, S.C. , Seed, B.V., 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. (doi:10.1111/j.1460-9568.2008.06415.x)

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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:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:McCulloch, Mrs Maj-Lis and Barnett, Professor Susan and McLaughlin, Dr Mark and Griffiths, Prof Ian
Authors: Thomson, C.E., McCulloch, M., Sorenson, A., Barnett, S.C., Seed, B.V., Griffiths, I.R., and McLaughlin, M.
College/School:College of Medical Veterinary and Life Sciences > School of Veterinary Medicine
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:European Journal of Neuroscience
Journal Abbr.:Europ. J. Neurosci.
Publisher:Wiley
ISSN:0953-816X
ISSN (Online):1460-9568
Published Online:11 September 2008
Copyright Holders:Copyright © 2008 The Authors
First Published:First published in European Journal of Neuroscience 28(8):1518-1535
Publisher Policy:Reproduced under a Creative Commons License

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