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Human Ntera2 cells as a predictive in vitro test system for developmental neurotoxicity

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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.

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Acknowledgments

This work was supported by the German Ministry of Education and Research (BMBF Grants 0313925D and 0315522D).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm 15/102, 30173, Hannover, Germany

    Michael Stern, Andrea Gierse, Saime Tan & Gerd Bicker

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  1. Michael Stern
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  2. Andrea Gierse
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  4. Gerd Bicker
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Correspondence to Michael Stern or Gerd Bicker.

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Stern, M., Gierse, A., Tan, S. et al. Human Ntera2 cells as a predictive in vitro test system for developmental neurotoxicity. Arch Toxicol 88, 127–136 (2014). https://doi.org/10.1007/s00204-013-1098-1

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