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.

اظهر المزيد [+]

الكلمات المفتاحية الخاصة بالمكنز الزراعي (أجروفوك)

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

المعلومات البيبليوغرافية

نُشرت في

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

المجلد 33 الإصدار 5 ترقيم الصفحات 1135 - 1146

الناشر Pathotox

الرقم التسلسلي المعياري الدولي (ردمد) 0161-813X

مواضيع أخرى

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

اللغة

إنجليزي

النوع

Journal Article; Text

في أجريس منذ: 2025-02-03

تاريخ التعديل: 2025-10-25

نوع الملف: MODS

مزود البيانات

تم تزويد هذا السجل من قبل German Federal Institute for Risk Assessment

اكتشف مجموعة مزود البيانات هذا في أجريس

الروابط

DOI https://www.openagrar.de/receive/bimport_mods_00001335

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

2012

الكلمات المفتاحية الخاصة بالمكنز الزراعي (أجروفوك)

المعلومات البيبليوغرافية