A culture model for neurite regeneration of human spinal cord neurons

J Neurosci Methods. 2011 Oct 15;201(2):346-54. doi: 10.1016/j.jneumeth.2011.08.024. Epub 2011 Aug 19.

Abstract

Effective therapeutic interventions for injuries of the central nervous system such as spinal cord injury are still unavailable, having a great impact on the quality of life of victims and their families, as well as high costs in medical care. Animal models of spinal cord injury are costly, time-consuming and labor-intensive, making them unsuitable for screening large numbers of experimental conditions. Thus, culture models that recapitulate key aspects of neuronal changes in central nervous system injuries are needed to gain further understanding of the pathological and regenerative mechanisms involved, as well as to accelerate the screening of potential therapeutic agents. In this study we differentiated adherent cultures of dissociated human fetal spinal cord neural precursors into postmitotic neurons which we could then detach from culture plates and successfully freeze down in a viable state. When replated in neuronal medium without neurodifferentiating factors, these ready-to-use human spinal cord neurons remained viable, postmitotic and regenerated neurites in a cell density-dependent manner. Insulin-like growth factor 1 and growth hormone had no effect on neurite regeneration while brain-derived neurotrophic factor increased both the number of cells with neurites as well as the average neurite length. Our model can be applied to investigate factors involved in neuroregeneration of the human spinal cord and since adherent dissociated cell cultures are used, this system has significant potential as a screening platform for therapeutic agents to treat spinal cord injury.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Drug Evaluation, Preclinical / methods
  • Fetus / cytology
  • Humans
  • Nerve Growth Factors / pharmacology
  • Nerve Growth Factors / therapeutic use
  • Nerve Regeneration / drug effects
  • Nerve Regeneration / physiology*
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / physiology
  • Neurites / drug effects
  • Neurites / physiology*
  • Spheroids, Cellular
  • Spinal Cord / cytology*
  • Spinal Cord / physiology*

Substances

  • Nerve Growth Factors