Neural differentiation of mouse embryonic stem cells as a tool to assess developmental neurotoxicity in vitro

Visan,A.; Hayess,K.; Sittner,D.; Pohl,E.E.; Riebeling,C.; Slawik,B.; Gulich,K.; Oelgeschläger,M.; Luch,A.; Seiler,A.E.M.

Neural differentiation of mouse embryonic stem cells as a tool to assess developmental neurotoxicity in vitro

2012

Mouse embryonic stem cells (mESCs) represent an attractive cellular system for in vitro studies in developmental biology as well as toxicology because of their potential to differentiate into all fetal cell lineages. The present study aims to establish an in vitro system for developmental neurotoxicity testing employing mESCs. We developed a robust and reproducible protocol for fast and efficient differentiation of the mESC line D3 into neural cells, optimized with regard to chemical testing. Morphological examination and immunocytochemical staining confirmed the presence of different neural cell types, including neural progenitors, neurons, astrocytes, oligodendrocytes, and radial glial cells. Neurons derived from D3 cells expressed the synaptic proteins PSD95 and synaptophysin, and the neurotransmitters serotonin and +¦-aminobutyric acid. Calcium ion imaging revealed the presence of functionally active glutamate and dopamine receptors. In addition, flow cytometry analysis of the neuron-specific marker protein MAP2 on day 12 after induction of differentiation demonstrated a concentration dependent effect of the neurodevelopmental toxicants methylmercury chloride, chlorpyrifos, and lead acetate on neuronal differentiation. The current study shows that D3 mESCs differentiate efficiently into neural cells involving a neurosphere-like state and that this system is suitable to detect adverse effects of neurodevelopmental toxicants. Therefore, we propose that the protocol for differentiation of mESCs into neural cells described here could constitute one component of an in vitro testing strategy for developmental neurotoxicity.

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AGROVOC Keywords

alternative methods analysis animal animals antibodies assessment behaviour berlin biology blood brain calcium cell differentiation cell structure cells central nervous system chemical analysis chemicals children chlorpyrifos cns concentration culture media cysteine development dna dopamine effects evaluation experiments fibroblasts flow cytometry gene gene expression germany groups health hormone humans identification immunocytochemistry in vitro infant lead lesions methods mice microrna migration model models myocardium nervous system neurotoxicity nutrition organization pathology pesticide pesticides poisoning polychlorinated biphenyls population prediction protein proteins proteomics rat reproducibility risk risk assessment science screening serotonin staining state strategies system systems technology teratogenicity testing tests therapy tissue toxicity toxicology usa

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Published in

Neurotoxicology (Park Forest South, Ill) | Neurotoxicology (Park For South) | Neurotoxicology

Volume 33 Issue 5 Pagination 1135 - 1146

Publisher Pathotox

ISSN 0161-813X

Other Subjects

Blood brain barrier; Prediction model; Proteomic; Ecvam; Thyroid-hormone; Phase; Parkinson's disease; Target; Represent; Nonhuman; Astrocytes; Embryotoxicity; In-vitro; Cultures; Validation; Markers; Guanylate kinase; Process optimization; Polychlorinated-biphenyls; N-methylaspartate; Stem cells; High throughput screening; Microtubule-associated proteins; Testing strategies; Organometallic compounds; In vitro testing strategy; Pathway; Time factors; Adverse; Fate; Calcium ion; Embryo; Technologies; Laminin; Neurite outgrowth; Reach; Cell survival; Membrane-protein; Embryotoxicity test; Animal-model; International; Region; Protein analysis; Alternative; Embryonic stem cells; Tubulin; Human; Conversion; Central-nervous-system; Developmental neurotoxicity; Neural stem cell; Glial fibrillary acidic protein; Neural differentiation; Est; Vitro; Potential; In vitro study; Developmental toxicity; Disease; Oligodendroglia; Report; 2'; 3'-cyclic-nucleotide phosphodiesterases; Vitro embryotoxicity tests; Human exposure; Glutamate receptor; Insights; Pattern; Activation; Differentiation; Alternatives; Mechanism; Line; Proliferation; Dopamine receptor; Culture; Detect; Strategy; Effect; Young adult; Bfr; Risk-assessment; Adverse effects; Biphenyls; Concentration response; Comparison; Sodium glutamate; Pathways; Biomarker; Testing strategy; Hazard assessment; Expression; Toxicity testing; Compound; Cell type; Compounds; Priority journal; Studies; Acid; Targets; Embryonic stem cell; Long-term; Validation study; Lead acetate; Induction; Glia cell; Exposure; Microtubule associated protein 2; 4 aminobutyric acid; Thyroid hormone receptors; Assay; In vitro system; Map; Alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid; Chemical; Article; Prediction models; Predict; Cell; Cell lineage; Protein expression; 37; In vitro developmental neurotoxicity testing; In vitro testing; Component; Animal cell; Rat brain; Animal model; Animal experiments; Applicability domain; Methylmercury compounds; Neuronal differentiation; Cell proliferation; Study; Method; Yolk; Antibody; Animal experiment; Reference; Postsynaptic density protein 95; Gene-expression; Alternative method; Controlled study; Methylmercuric chloride; Different regions; Animal-experiments; Mouse; Neurons; Progenitors; Synaptophysin; Organophosphate; Interpretation; Astrocyte; Hazard; Cell-line; Nerve cell differentiation; Glutamate sodium; Membrane proteins; In-utero; Immunofluorescence microscopy; Embryonic stem cell test; Protocol; Review; Tyrosine 3-monooxygenase; Induce; Cell line; Adult

Language

English

Type

Text; Journal Article

In AGRIS since: 2025-02-03

Modification date: 2025-07-17

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Neural differentiation of mouse embryonic stem cells as a tool to assess developmental neurotoxicity in vitro

2012

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