Neuron-like cells generated from human umbilical cord lining-derived Mesenchymal stem cells as a new in vitro model for neuronal toxicity screening: Using magnetite nanoparticles as an example

Uliana De Simone, Arsenio Spinillo, Francesca Caloni, Laura Gribaldo, Teresa Coccini

Research output: Contribution to journalArticle

Abstract

The wide employment of iron nanoparticles in environmental and occupational settings underlines their potential to enter the brain. Human cell-based systems are recommended as relevant models to reduce uncertainty and to improve prediction of human toxicity. This study aimed at demonstrating the in vitro differentiation of the human umbilical cord lining-derived-mesenchymal stem cells (hCL-MSCs) into neuron-like cells (hNLCs) and the benefit of using them as an ideal primary cell source of human origin for the neuronal toxicity of Fe3O4 NPs (magnetite-nanoparticles). Neuron-like phenotype was confirmed by: live morphology; Nissl body staining; protein expression of different neuronal-specific markers (immunofluorescent staining), at different maturation stages (i.e., day-3-early and day-8-full differentiated), namely β-tubulin III, MAP-2, enolase (NSE), glial protein, and almost no nestin and SOX-2 expression. Synaptic makers (SYN, GAP43, and PSD95) were also expressed. Fe3 O4 NPs determined a concentration-and time-dependent reduction of hNLCs viability (by ATP and the Trypan Blue test). Cell density decreased (20–50%) and apoptotic effects were detected at ≥10 µg/mL in both types of differentiated hNLCs. Three-day-differentiated hNLCs were more susceptible (toxicity appeared early and lasted for up to 48 h) than 8-day-differentiated cells (delayed effects). The study demonstrated that (i) hCL-MSCs easily differentiated into neuronal-like cells; (ii) the hNCLs susceptibility to Fe3 O4 NPs; and (iii) human primary cultures of neurons are new in vitro model for NP evaluation.

Original languageEnglish
Article number271
JournalInternational Journal of Molecular Sciences
Volume21
Issue number1
DOIs
Publication statusPublished - Jan 1 2020

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Keywords

  • Alternative methods
  • Cell-based assay
  • Environmental toxicology
  • Fe O nanoparticles
  • Human primary cell culture
  • Predictive nanotoxicology
  • Safety assessment
  • Toxicity-testing strategies

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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