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Article

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

1
Laboratory of Clinical & Experimental Toxicology, Toxicology Unit, ICS Maugeri SpA-Benefit Corporation, IRCCS Pavia, Via Maugeri 10, 27100 Pavia, Italy
2
Department of Obstetrics and Gynecology, Fondazione IRCCS Policlinico San Matteo and University of Pavia, 27100 Pavia, Italy
3
Dipartimento di Medicina Veterinaria (DIMEVET), Università degli Studi di Milano, 20133 Milano, Italy
4
Chemical Safety and Alternative Methods Unit, Directorate F—Health, Consumers and Reference Materials, Directorate General Joint Research Centre, European Commission, 21027 Ispra, Italy
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(1), 271; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010271
Received: 27 November 2019 / Revised: 20 December 2019 / Accepted: 29 December 2019 / Published: 31 December 2019
(This article belongs to the Special Issue Nanotoxicology and Nanosafety 2.0)
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 Fe3O4NPs (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. Fe3O4NPs 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 Fe3O4NPs; and (iii) human primary cultures of neurons are new in vitro model for NP evaluation. View Full-Text
Keywords: Fe3O4 nanoparticles; environmental toxicology; alternative methods; safety assessment; cell-based assay; toxicity-testing strategies; human primary cell culture; predictive nanotoxicology Fe3O4 nanoparticles; environmental toxicology; alternative methods; safety assessment; cell-based assay; toxicity-testing strategies; human primary cell culture; predictive nanotoxicology
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MDPI and ACS Style

De Simone, U.; Spinillo, A.; Caloni, F.; Gribaldo, L.; Coccini, T. 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. Int. J. Mol. Sci. 2020, 21, 271. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010271

AMA Style

De Simone U, Spinillo A, Caloni F, Gribaldo L, Coccini T. 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. International Journal of Molecular Sciences. 2020; 21(1):271. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010271

Chicago/Turabian Style

De Simone, Uliana, Arsenio Spinillo, Francesca Caloni, Laura Gribaldo, and Teresa Coccini. 2020. "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" International Journal of Molecular Sciences 21, no. 1: 271. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010271

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