Methods for Culturing Primary Sympathetic Neurons and for Determining Neuronal Viability

  • Jonathan Whitfield
  • Stephen J. Neame
  • Jonathan Ham
Part of the Methods in Molecular Biology book series (MIMB, volume 282)


Developing nerve growth factor (NGF)-dependent sympathetic neurons are one of the best-studied in vitro models of neuronal apoptosis and have been used to identify key components of the neuronal cell death pathway. This chapter describes how to prepare purified cultures of primary sympathetic neurons and how to induce apoptosis by NGF deprivation. In addition, a simple method for measuring neuronal viability based on the live/dead assay is also described. This can be used for assessing the effect of small molecule inhibitors of protein kinases, caspases and other enzymes, on NGF withdrawal-induced death.

Key Words

Sympathetic neuron nerve growth factor developmental neuronal death apoptosis live/dead cell viability assay 


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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Jonathan Whitfield
    • 1
  • Stephen J. Neame
    • 1
  • Jonathan Ham
    • 2
  1. 1.Eisai London Research LaboratoriesUniversity College LondonLondonUK
  2. 2.Molecular Hematology and Cancer Biology Unit, Institute of Child HealthUniversity College LondonLondonUK

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