Goette, Marjo, Hofmann, Gabriele, Gallani, Anne-Isabelle, Glickman, Fraser, Wishart, William L. and Gabriel, Daniela (2010) An imaging assay to analyze primary neurons for cellular neurotoxicity. Journal of Neuroscience Methods, 192 (1). pp. 7-16.

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.

Item Type:	Article
Related URLs:	http://www.ncbi.nlm.nih.gov/sites/entrez
Additional Information:	author can archive post-print (ie final draft post-refereeing); Publisher's version/PDF cannot be used
Keywords:	high-content analysis, neurite outgrowth, neurotoxicity, cerebellar granule neurons
Related URLs:	http://www.ncbi.nlm.nih.gov/sites/entrez
Date Deposited:	13 Oct 2015 13:16
Last Modified:	13 Oct 2015 13:16
URI:	https://oak.novartis.com/id/eprint/2700