An in vitro model of adult mammalian nerve repair

Exp Neurol. 2010 May;223(1):112-8. doi: 10.1016/j.expneurol.2009.05.022. Epub 2009 May 21.


The role of pathway-derived growth factors in the support of peripheral axon regeneration remains elusive. Few appropriate knock-out mice are available, and gene silencing techniques are rarely 100% effective. To overcome these difficulties, we have developed an in vitro organotypic co-culture system that accurately models peripheral nerve repair in the adult mammal. Spinal cord sections from P4 mice that express YFP in their neurons are used to innervate segments of P4 peripheral nerve. This reconstructed ventral root is then transected and joined to a nerve graft. Growth of axons across the nerve repair and into the graft can be imaged repeatedly with fluorescence microscopy to define regeneration speed, and parent neurons can be labeled in retrograde fashion to identify contributing neurons. Nerve graft harvested from adult mice remains viable in culture by both morphologic and functional criteria. Motoneurons are supported with GDNF for the first week in culture, after which they survive axotomy, and are thus functionally adult. This platform can be modified by using motoneurons from any genetically modified mouse that can be bred to express XFP, by harvesting nerve graft from any source, or by treating the culture systemically with antibodies, growth factors, or pathway inhibitors. The regeneration environment is controlled to a degree not possible in vivo, and the use of experimental animals is reduced substantially. The flexibility and control offered by this technique should thus make it a useful tool for the study of regeneration biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Coculture Techniques / methods
  • Dextrans
  • Glial Cell Line-Derived Neurotrophic Factor / pharmacology
  • Insulin-Like Growth Factor I / pharmacology
  • Luminescent Proteins / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Electron, Transmission / methods
  • Models, Animal*
  • Motor Neurons / drug effects
  • Motor Neurons / physiology*
  • Nerve Growth Factor / pharmacology
  • Nerve Regeneration / drug effects
  • Nerve Regeneration / genetics
  • Nerve Regeneration / physiology*
  • Organ Culture Techniques / methods
  • Peripheral Nerves / physiology
  • Peripheral Nerves / ultrastructure
  • Rhodamines
  • Spinal Cord / cytology


  • Dextrans
  • Fluoro-Ruby
  • Glial Cell Line-Derived Neurotrophic Factor
  • Luminescent Proteins
  • Rhodamines
  • Insulin-Like Growth Factor I
  • Nerve Growth Factor