UNIT 11.16 Morphological Assessment of Neurite Outgrowth in Hippocampal Neuron-Astrocyte Co-Cultures

  1. Gennaro Giordano1,
  2. Lucio G. Costa1,2

Published Online: 1 MAY 2012

DOI: 10.1002/0471140856.tx1116s52

Current Protocols in Toxicology

Current Protocols in Toxicology

How to Cite

Giordano, G. and Costa, L. G. 2012. Morphological Assessment of Neurite Outgrowth in Hippocampal Neuron-Astrocyte Co-Cultures. Current Protocols in Toxicology. 52:11.16:11.16.1–11.16.26.

Author Information

  1. 1

    Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington

  2. 2

    Department of Human Anatomy, Pharmacology and Forensic Science, University of Parma Medical School, Parma, Italy

Publication History

  1. Published Online: 1 MAY 2012
  2. Published Print: MAY 2012


Neurite outgrowth is a fundamental event in brain development, as well as in regeneration of damaged neurons. Astrocytes play a major role in neuritogenesis, by expressing and releasing factors that facilitate neurite outgrowth, such as extracellular matrix proteins, and factors that can inhibit neuritogenesis, such as the chondroitin sulfate proteoglycan neurocan. In this unit we describe a noncontact co-culture system of hippocampal neurons and cortical (or hippocampal) astrocytes for measurement of neurite outgrowth. Hippocampal pyramidal neurons are plated on glass coverslips, which are inverted onto an astrocyte feeder layer, allowing exposure of neurons to astrocyte-derived factors without direct contact between these two cell types. After co-culture, neurons are stained and photographed, and processes are assessed morphologically using Metamorph software. This method allows exposing astrocytes to various agents before co-culture in order to assess how these exposures may influence the ability of astrocytes to foster neurite outgrowth. Curr. Protoc. Toxicol. 52:11.16.1-11.16.26. © 2012 by John Wiley & Sons, Inc.


  • pyramidal hippocampal neurons;
  • astrocytes;
  • co-culture;
  • neurite outgrowth