A model for ex vivo spinal cord segment culture--a tool for analysis of injury repair strategies

J Neurosci Methods. 2010 Sep 30;192(1):49-57. doi: 10.1016/j.jneumeth.2010.07.012. Epub 2010 Jul 21.

Abstract

Most spinal cord injury research is undertaken using in vivo animal models but the extensive care associated with spinalized animals, inherent variability between animals, and complex surgeries makes alternative models especially valuable. Here we present a novel ex vivo model that enables culture of intact post-natal spinal cord segments for up to five days and the assessment of peripheral nerve grafting repair, enhanced with connexin43 antisense oligodeoxynucleotides (Cx43 AsODN), in this model. Down-regulating Cx43 expression with Cx43 AsODN in cultured spinal cord segments prevents cell death and inhibits inflammation spreading from the site of injury to neighbouring tissue, hence maintaining culture viability. Reduction in segment swelling and improvement in neuron survival were evident after Cx43 AsODN treatment. Furthermore, the combination of Cx43 AsODN with peripheral nerve graft implants into cultured spinal cords promoted axon sprouting from the spinal cord into the peripheral nerve graft. This ex vivo spinal cord segment culture model provides a valuable addition to tools currently available for spinal cord injury research.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Axons / drug effects
  • Axons / physiology
  • Cell Survival / drug effects
  • Connexin 43 / metabolism
  • Dose-Response Relationship, Drug
  • Edema / drug therapy
  • Edema / pathology
  • Gene Expression Regulation / drug effects
  • Nerve Regeneration / drug effects
  • Nerve Regeneration / physiology*
  • Neurites / drug effects
  • Neurofilament Proteins / metabolism
  • Neurons / cytology
  • Neurons / drug effects*
  • Oligodeoxyribonucleotides, Antisense / pharmacology
  • Organ Culture Techniques
  • Rats
  • Sciatic Nerve / cytology
  • Sciatic Nerve / transplantation*
  • Spinal Cord / cytology
  • Spinal Cord / physiology*
  • Spinal Cord / surgery*

Substances

  • Connexin 43
  • Neurofilament Proteins
  • Oligodeoxyribonucleotides, Antisense
  • neurofilament protein H