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J Neurosci Methods. 2010 Sep 30;192(1):7-16. doi: 10.1016/j.jneumeth.2010.07.003. Epub 2010 Jul 8.

An imaging assay to analyze primary neurons for cellular neurotoxicity.

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1
Novartis Institutes for BioMedical Research, Forum 1, Novartis Campus, CH-4002 Basle, Switzerland. marjo.goette@novartis.com

Abstract

The development of high-content screening technologies including automated immunostaining, automated image acquisition and automated image analysis have enabled higher throughput of cellular imaging-based assays. Here we used high-content imaging to thoroughly characterize the cultures of primary rat cerebellar granule neurons (CGNs). We describe procedures to isolate and cultivate the CGNs in 96-well and 384-well format, as well as a procedure to freeze and thaw the CGNs. These methods allow the use of CGNs in 96-well format analyzing 2500 samples per experiment using freshly isolated cells. Down-scaling to 384-well format and freezing and thawing of the CGNs allow even higher throughput. A cellular assay with rat CGN cultures was established to study the neurotoxicity of compounds in order to filter out toxic compounds at an early phase of drug development. The imaging-based toxicity assay was able to reveal adverse effects of compounds on primary neurons which were not detected in neuroblastoma or other cell lines tested.

PMID:
20620166
DOI:
10.1016/j.jneumeth.2010.07.003
[Indexed for MEDLINE]
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