Differentiating human NT2/D1 neurospheres as a versatile in vitro 3D model system for developmental neurotoxicity testing

Eric J. Hill, Elizabeth K. Woehrling, Mark Prince, Mike D. Coleman

Research output: Contribution to journalArticle

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.
LanguageEnglish
Pages243-250
Number of pages8
JournalToxicology
Volume249
Issue number2-3
Early online date30 May 2008
DOIs
Publication statusPublished - 30 Jul 2008

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Hydroxyurea
Acrylamide
Clone Cells
Neurogenesis
Valproic Acid
Lithium
Cell Survival
Testing
Clone cells
Cells
Teratogens
Proteins
Neurotoxins
Neurology
Central Nervous System
Health
In Vitro Techniques

Keywords

  • acrylamides
  • Western blotting
  • cell differentiation
  • tumor cell line
  • cell survival
  • densitometry
  • gene expression
  • humans
  • hydroxyurea
  • computer-assisted image processing
  • immunohistochemistry
  • lithium chloride
  • confocal microscopy
  • neurological models
  • nervous system diseases
  • neurons
  • reverse transcriptase polymerase chain rReaction
  • teratogens
  • tretinoin
  • valproic acid

Cite this

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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.",
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Differentiating human NT2/D1 neurospheres as a versatile in vitro 3D model system for developmental neurotoxicity testing. / Hill, Eric J.; Woehrling, Elizabeth K.; Prince, Mark; Coleman, Mike D.

In: Toxicology, Vol. 249, No. 2-3, 30.07.2008, p. 243-250.

Research output: Contribution to journalArticle

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