title: Differentiating human NT2/D1 neurospheres as a versatile in vitro 3D model system for developmental neurotoxicity testing.
abstract: Developmental neurotoxicity is a major issue in human health and may have lasting neurological implications. In this preliminary study we exposed differentiating Ntera2/clone D1 (NT2/D1) cell neurospheres to known human teratogens classed as non-embryotoxic (acrylamide), weakly embryotoxic (lithium, valproic acid) and strongly embryotoxic (hydroxyurea) as listed by European Centre for the Validation of Alternative Methods (ECVAM) and examined endpoints of cell viability and neuronal protein marker expression specific to the central nervous system, to identify developmental neurotoxins. Following induction of neuronal differentiation, valproic acid had the most significant effect on neurogenesis, in terms of reduced viability and decreased neuronal markers. Lithium had least effect on viability and did not significantly alter the expression of neuronal markers. Hydroxyurea significantly reduced cell viability but did not affect neuronal protein marker expression. Acrylamide reduced neurosphere viability but did not affect neuronal protein marker expression. Overall, this NT2/D1-based neurosphere model of neurogenesis, may provide the basis for a model of developmental neurotoxicity in vitro.
authors.affiliation: School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, UK. hillej@aston.ac.uk
authors.forename: E J
authors.initials: EJ
authors.lastname: Hill
authors.forename: E K
authors.initials: EK
authors.lastname: Woehrling
authors.forename: M
authors.initials: M
authors.lastname: Prince
authors.forename: M D
authors.initials: MD
authors.lastname: Coleman
mesh_terms: D000178:Acrylamides / Q000633:toxicity; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003720:Densitometry; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D006918:Hydroxyurea / Q000633:toxicity; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D018021:Lithium Chloride / Q000633:toxicity; D018613:Microscopy, Confocal; D008959:Models, Neurological; D009422:Nervous System Diseases / Q000139:chemically induced* / Q000473:pathology*; D009474:Neurons / Q000187:drug effects / Q000473:pathology / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013723:Teratogens / Q000633:toxicity*; D014212:Tretinoin / Q000633:toxicity; D014635:Valproic Acid / Q000633:toxicity
chemical_list: D000178:Acrylamides; D013723:Teratogens; D014212:Tretinoin; D014635:Valproic Acid; D018021:Lithium Chloride; D006918:Hydroxyurea