Assessment of toxicity responses in primary cortical neurons

Summary: An assessment and prediction of drug-induced neuroprotection and toxicity in rat primary cortical neurons.

Model Description

An assessment and prediction of study compound-induced neuroprotection / toxicity on the rat primary cortical neurons in vitro. 

Species Rat primary cortical neurons 
Cell viability assessment Rat cortical neurons are grown on 96-well plates. The cells can be exposed to testing molecules or formulations for selected times (e.g. 4h, 24h). Cytotoxicity is measured using resazurin reduction assay. A cell permeable redox indicator resazurin (7-Hydroxy-3H-phenoxazin-3-one-10-oxide) is used to measure oxidation-reduction reactions in viable cells. The non-fluorescent resazurin is irreversibly reduced by metabolically active cells to highly fluorescent metabolite resorufin (7-Hydroxy-3H-phenoxazin-3-one). Resorufin formation is measured by using Cytation 3 multi-mode reader (BioTek Instruments, Winooski, VT, USA).  The quantity of resorufin produced is proportional to the number of viable cells. 
Oxidative stress inducer tert-butyl hydroperoxide (tBHP) at concentrations 0-40 mM 
Positive control (±)-6-Hydroxy-2,5,7,8- tetramethylchromane-2-carboxylic acid (Trolox) 1000 mM 
Read-outs Cell viability (%)
IC50 value (50% cell viability) for positive control
IC50 value is calculated for study compound if applicable  

Outcomes and Read-Outs 

Fig. 1. Immunostaining of the rat primary cortical neurons (in red) counterstained with cellular nuclear marker DAPI (in blue).
Fig. 1. Immunostaining of the rat primary cortical neurons (in red) counterstained with cellular nuclear marker DAPI (in blue). 

Authors' picture