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Quantitative Assessment of Neurite Outgrowth in PC12 Cells
Citation:
HARRILL, J. AND W. R. MUNDY. Quantitative Assessment of Neurite Outgrowth in PC12 Cells. Chapter 23, L. Costa, G. Giordano, M.Guizzetti (ed.), In Vitro Neurotoxicology: Methods and Protocols; Methods in Molecular Biology Series. Springer, New York, NY, 1:331-348, (2011).
Impact/Purpose:
This chapter provides, in detail, two methods to quantify the number of neurite bearing cells and neurite length in PC12 cells.The first method is based on the imaging of live cells using phase contrast or interference contrast microscopy, followed by manual tracing of neurite length. The second method uses a high content image analysis system that automates the acquisition and analysis of neurite outgrowth in cells that have been immunocytochemically labeled with a fluorescent tag.
Description:
In vitro test methods can provide a rapid approach for the screening of large numbers of chemicals for their potential to produce toxicity. In order to identify potential developmental neurotoxicants, assessment of critical neurodevelopmental processes such as neuronal differentiation and growth has been proposed. PC12 cells have been widely used to study the neurotrophic factor-induced signaling pathways that control differentiation, and as in vitro models to detect the effect of chemicals on neurite outgrowth. Upon exposure to nerve growth factor (NGF), PC12 cells cease to proliferate, extend multiple neurites, and acquire the properties of sympathetic neurons. Measurement of the number and length ofneurites during exposure to NGF provides a quantitative assessment of neuronal differentiation and growth. Differentiation and neurite outgrowth can be measured using simple contrast microscopy in live cells, or using automated imaging systems in cells prepared with immunocytochemistry