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Example sentences for: wst-
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The cytotoxicity of domoic acid to neuronal tissue in vitro has been investigated over a wide range of concentrations, namely at 10 μM with primary cultures of cerebellar neurons [ 30 31 ] as well as hippocampal slices [ 33 ] , at 100 μM with cultured prenatal rat hippocampal neurons [ 29 ] and at 20 mM with rat lumbar spinal interneurons [ 28 ] . In order to determine a concentration of domoic acid for our in vitro studies that was toxic to cerebellar granule cells after a short-term exposure in our experimental conditions, we completed a viability study using the WST-1 method.
Domoic acid has been reported to be neurotoxic to neuronal tissue in vitro in concentrations ranging from 10 μM to 20 mM [ 28 29 30 31 32 33 ] . In order to determine a concentration of domoic acid that was neurotoxic in vitro during a 6 hour exposure, we investigated the concentration-dependent effect of domoic acid on cerebellar granule neurons using the WST-1 viability assay.
Plates were incubated in a humidified 5% CO 2 incubator at 36°C and the reduction of the WST-1 reagent to formazan was measured repeatedly after the addition of the WST-1 reagent, at either 415 nM (neurons) or 450 nM (microglia), with a microtiter plate (ELISA) reader.
Cells were then returned into the incubator for 40-45 hrs before the addition of 1/10 vol of WST-1.
A colorimetric assay for the quantitation of cell viability, based on the cleavage of the tetrazolium salt WST-1 (4-[3-(4-Iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]1,3-benzene disulfonate) (slightly red) to formazan (red) by mitochondrial dehydrogenases in viable cells was utilized for these studies.
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