Stem Cell Research (2017-12-01)

Comparative performance analysis of human iPSC-derived and primary neural progenitor cells (NPC) grown as neurospheres in vitro

  • Maxi Hofrichter,
  • Laura Nimtz,
  • Julia Tigges,
  • Yaschar Kabiri,
  • Friederike Schröter,
  • Brigitte Royer-Pokora,
  • Barbara Hildebrandt,
  • Martin Schmuck,
  • Alexey Epanchintsev,
  • Stephan Theiss,
  • James Adjaye,
  • Jean-Marc Egly,
  • Jean Krutmann,
  • Ellen Fritsche

DOI
https://doi.org/10.1016/j.scr.2017.10.013
Journal volume & issue
Vol. 25, no. C
pp. 72 – 82

Abstract

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Developmental neurotoxicity (DNT) testing performed in rats is resource-intensive (costs, time, animals) and bears the issue of species extrapolation. Thus, reliable alternative human-based approaches are needed for predicting neurodevelopmental toxicity. Human induced pluripotent stem cells (hiPSCs) represent a basis for an alternative method possibly being part of an alternative DNT testing strategy. Here, we compared two hiPSC neural induction protocols resulting in 3D neurospheres: one using noggin and one cultivating cells in neural induction medium (NIM protocol). Performance of Nestin+/SOX2+ hiPSC-derived neural progenitor cells (NPCs) was compared to primary human NPCs. Generally, primary hNPCs first differentiate into Nestin+ and/or GFAP+ radial glia-like cells, while the hiPSC-derived NPCs (hiPSC-NPC) first differentiate into βIII-Tubulin+ neurons suggesting an earlier developmental stage of hiPSC-NPC. In the ‘Neurosphere Assay’, NIM generated hiPSC-NPC produced neurons with higher performance than with the noggin protocol. After long-term differentiation, hiPSC-NPC form neuronal networks, which become electrically active on microelectrode arrays after 85 days. Finally, methylmercury chloride inhibits hiPSC-NPC and hNPC migration with similar potencies. hiPSC-NPCs-derived neurospheres seem to be useful for DNT evaluation representing early neural development in vitro. More system characterization by compound testing is needed to gain higher confidence in this method.

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