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

C.E. Thomson, M. McCulloch, Annette Sorensen, S.C. Barnett, B.V. Seed, I.R. Griffiths, M. McLaughlin

Research output: Contribution to journalArticle

35 Citations (Scopus)

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.
LanguageEnglish
Pages1518-1535
Number of pages17
JournalEuropean Journal of Neuroscience
Volume28
Issue number8
DOIs
Publication statusPublished - Oct 2008

Fingerprint

Myelin Sheath
Spinal Cord
Central Nervous System
Axons
Synapses
Research
Ranvier's Nodes
Myelin Proteins
Systems Biology
Central Nervous System Diseases
Oligodendroglia
Microscopy
In Vitro Techniques
Pathology
Cytokines
Brain
Serum

Keywords

  • cytokines
  • MAG
  • multiple sclerosis
  • PLP-DM20
  • western blotting
  • spinal cord

Cite this

Thomson, C.E. ; McCulloch, M. ; Sorensen, Annette ; Barnett, S.C. ; Seed, B.V. ; Griffiths, I.R. ; McLaughlin, M. / Myelinated, synapsing cultures of murine spinal cord - validation as an in vitro model of the central nervous system. In: European Journal of Neuroscience. 2008 ; Vol. 28, No. 8. pp. 1518-1535.
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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.; McCulloch, M.; Sorensen, Annette; Barnett, S.C.; Seed, B.V.; Griffiths, I.R.; McLaughlin, M.

In: European Journal of Neuroscience, Vol. 28, No. 8, 10.2008, p. 1518-1535.

Research output: Contribution to journalArticle

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T1 - Myelinated, synapsing cultures of murine spinal cord - validation as an in vitro model of the central nervous system

AU - Thomson, C.E.

AU - McCulloch, M.

AU - Sorensen, Annette

AU - Barnett, S.C.

AU - Seed, B.V.

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AU - McLaughlin, M.

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