title: Human Ntera2 cells as a predictive in vitro test system for developmental neurotoxicity.
abstract: Developmental neurotoxicity (DNT) of environmental chemicals is a serious threat to human health. Current DNT testing guidelines propose investigations in rodents, which require large numbers of animals. With regard to the "3Rs" (reduction, replacement, and refinement) of animal testing, alternative testing strategies are needed in order to refine and reduce animal experiments and allow faster and less expensive screening. The goal of this study was to establish components for a human cell-based test system to assess DNT potential of chemicals at an early stage of brain development. A human neural precursor cell line should be tested for suitability for semi-automated high-throughput DNT screening. We established assays suitable for detecting disturbances in two basic processes of brain development in 96-well scale: neuronal differentiation and migration using the human Ntera2 (NT2) cell line. We assessed the effects of four test compounds with well-established DNT potential in comparison with three compounds without specific DNT potential. We found that human NT2 cell cultures treated with the morphogen, retinoic acid, imitate neuronal differentiation, and migration in vitro. The developmental neurotoxicants methylmercury chloride, sodium arsenite, sodium valproate, and methylazoxymethanol significantly reduced the expression of the neuronal marker β-tubulin type III and decreased the migration distance in developing NT2 cells. Both endpoints, differentiation and migration, can be read out directly in a standard fluorescence plate reader, enabling high-throughput screening. We conclude that NT2 cell tests are likely to become valuable components of a human cell-based modular in vitro DNT test systems.
authors.affiliation: Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm 15/102, 30173, Hannover, Germany, Michael.stern@tiho-hannover.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Stern
authors.forename: Andrea
authors.initials: A
authors.lastname: Gierse
authors.forename: Saime
authors.initials: S
authors.lastname: Tan
authors.forename: Gerd
authors.initials: G
authors.lastname: Bicker
mesh_terms: D018053:Arsenites / Q000633:toxicity; D001921:Brain / Q000187:drug effects / Q000254:growth & development*; D002454:Cell Differentiation; D002460:Cell Line; D002465:Cell Movement; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008746:Methylazoxymethanol Acetate / Q000031:analogs & derivatives / Q000633:toxicity; D008767:Methylmercury Compounds / Q000633:toxicity; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D020258:Neurotoxicity Syndromes / Q000209:etiology*; D017670:Sodium Compounds / Q000633:toxicity; D018675:Toxicity Tests / Q000379:methods*; D014404:Tubulin / Q000378:metabolism; D014635:Valproic Acid / Q000633:toxicity
chemical_list: D018053:Arsenites; D008767:Methylmercury Compounds; D017670:Sodium Compounds; D014404:Tubulin; C017947:sodium arsenite; D008746:Methylazoxymethanol Acetate; D014635:Valproic Acid; C030667:methylazoxymethanol; C004925:methylmercuric chloride