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Rui Simoes. Picture.

Rui Simoes

Rui Simöes, Guest researcher

Rui Simoes. Picture.

Refinement of a differentiation protocol using neuroblastoma SH-SY5Y cells for use in neurotoxicology research

Author

  • Rui F Simões
  • Rafaela Ferrão
  • Margarida R Silva
  • Sonia L C Pinho
  • Lino Ferreira
  • Paulo J Oliveira
  • Teresa Cunha-Oliveira

Summary, in English

Since most models used to study neuronal dysfunction display disadvantages and ethical concerns, a fast and reproducible in vitro model to study mitochondria-related neurodegeneration is required. Here, we optimized and characterized a 3-day retinoic acid-based protocol to differentiate the SH-SY5Y cell line into a neuronal-like phenotype and investigated alterations in mitochondrial physiology and distribution. Differentiation was associated with p21-linked cell cycle arrest and an increase in cell mass and area, possibly associated with the development of neurite-like extensions. Notably, increased expression of mature neuronal markers (neuronal-specific nuclear protein, microtubule-associated protein 2, βIII tubulin and enolase 2) was observed in differentiated cells. Moreover, increased mitochondrial content and maximal area per cell suggests mitochondrial remodeling. To demonstrate that this model is appropriate to study mitochondrial dysfunction, cells were treated for 6 h with mitochondrial toxicants (rotenone, antimycin A, carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP) and 6-hydroxydopamine (6-OHDA)). Differentiated cells were more susceptible to increasing concentrations of FCCP, antimycin A, and rotenone, while 6-OHDA showed a distinct dose-dependent neurotoxicity pattern. Even though differentiated cells did not exhibit a fully mature/differentiated neuronal phenotype, the protocol developed can be used to study neurotoxicity processes, mitochondrial dynamics, and bioenergetic impairment, representing an alternative to study mitochondrial impairment-related pathologies in vitro.

Publishing year

2021

Language

English

Pages

1-13

Publication/Series

Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Volume

149

Document type

Journal article

Publisher

Elsevier

Keywords

  • Cell Differentiation
  • Cell Line, Tumor
  • Colorimetry
  • Humans
  • Microscopy/methods
  • Mitochondria/drug effects
  • Neuroblastoma
  • Neurons/drug effects
  • Neurotoxicity Syndromes/pathology
  • Rhodamines
  • Tretinoin/toxicity

Status

Published

ISBN/ISSN/Other

  • ISSN: 1873-6351