Establishment of a developmental neurotoxicity test by Sox1-GFP mouse embryonic stem cells

•Novel DNT Method was Built Employing Sox1-GFP cells.•Neural Differentiation Marker Sox1 Used to Evaluating Chemical Neurotoxicity.•Neuronal Differentiation of Sox1-GFP cells is Inhibited by the Neurotoxic Chemicals. Developmental toxicity tests have been generated by applying the embryonic stem cel...

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Published in:Reproductive toxicology (Elmsford, N.Y.) N.Y.), 2021-09, Vol.104, p.96-105
Main Authors: Park, Seon Mi, Jo, Na Rae, Lee, Bonn, Jung, Eui-Man, Lee, Sung Duck, Jeung, Eui-Bae
Format: Article
Language:English
Subjects:
46C cells
Animals
Cell Differentiation
Cell Line
Cell Survival
Cells, Cultured
DNT
Embryoid Bodies - metabolism
Green Fluorescent Proteins - metabolism
mESCs
Mice
Mouse Embryonic Stem Cells - metabolism
Neuroepithelial Cells
Neurons - drug effects
Neurons - metabolism
Neurotoxicity Syndromes
Sox1-GFP
SOXB1 Transcription Factors - metabolism
Toxicity Tests - methods
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Summary:•Novel DNT Method was Built Employing Sox1-GFP cells.•Neural Differentiation Marker Sox1 Used to Evaluating Chemical Neurotoxicity.•Neuronal Differentiation of Sox1-GFP cells is Inhibited by the Neurotoxic Chemicals. Developmental toxicity tests have been generated by applying the embryonic stem cell tests at the European Centre for the Validation of Alternative Methods, or by using the embryoid body test in our laboratory. This study was undertaken to explore novel developmental neurotoxicity (DNT) assay, using a Sox1-GFP cell line (mouse embryonic stem cells with an endogenous Sox1-GFP reporter). The expression of Sox1, a marker for neuroepithelial cells, is detected by green fluorescence, and the fluorescence intensity is a critical factor for achieving neuronal differentiation. Sox1-GFP cells cultured for 24 h were exposed to eleven neurotoxicants and four non-neurotoxicants. CCK-8 assays were performed to determine IC50 values after 48 h of chemical treatment. The fluorescence intensity of GFP was measured 4 days after treating the cells, and it was observed to decrease after exposure to neurotoxicants at higher concentrations, thereby indicating that the neuronal differentiation of Sox1-GFP cells is inhibited by the chemicals. Taken together, the results obtained in this study provide a model for DNT using embryonic stem cells, which may be applied to evaluate the toxicity of new chemicals or new drug candidates.
ISSN:0890-6238
1873-1708
DOI:10.1016/j.reprotox.2021.07.004