Development of an in vitro model of NanoNeurotoxicity using human 3D neural tissues: Functional assay using porous microelectrode arrays
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1
High School of Landscape Engineering and Architecture, University of Applied Sciences Western Switzerland, Switzerland
Given that many of the neurobehavioral/neurodegenerative diseases may be of environmental origin, further studies on the potential neurotoxic effects of NanoParticles (NPs), including the understanding the induction of various mechanisms leading to neuroinflammation and neurodegeneration associated with NPs are urgently needed. To answer this need, we propose to study t e biological and toxicological effects induced by NPs in the central nervous system by investigating the effects of the NPs on neuro-glial networks that may penetrate through to the brain parenchyma, using human in vitro models derived from stem-cells.
Neural stem cells derived from induced pluripotent stem cells (MTI-GlobalStem, ThermoFisher) are used to generate heterogeneous neural tissues which comprise different types of neurons and glial cells, as confirmed by histological studies. Neurospheres (NSs) are generated by aggregation of neuroprogenitors into 24 well plates under rotation. NPs are either added during the aggregation process or after the formation of the NSs. Au-NPs or Silica-FITC nanoparticles were tested in a first series of experiments. Fluorescent microscopy and transmission Electron Microscopy (TEM) were used to visualize the NPs within the nervous parenchyma. Electrophysiological recording of the activities of the neural networks were performed to assess the functionality of the neural tissue when exposed to the NPs.
Preliminary results (see figure) show as expected, that inclusion of Silica-FITC NPs (200nm; 10µg/ml) during the aggregation process are distributed within the entire NSs (A1 after 4 days) while when added after the formation of the NSs, they remain located in the out layers of the tissue (A2 after 4 days). In another series of experiments, Au-NP (50nm) were added during the NSs formation (D1=control, D2=0.5ppm, D3=5ppm and TEM insert). Electrophysiological recordings from those cultures after one month in culture show a clear decrease of the activities depending on the doses of the inclusions of NPs (Spikes: B1=control, B2=0.5ppm, B3=5ppm) Timestamps (C1=control, C2=0.5ppm, C3=5ppm).
Acknowledgements
The authors acknowledge funding from the NanoReg2 H2020 project, the Swiss Centre of Applied Human Toxicology (SCAHT) and the HES-SO and thank Jörg Huwyler - UniBasel, Jerome Rose - CEREGE, Iryna Nikonenko - UniGe, and Jerome Extermann – Hepia.
Keywords:
Porous polyimide microelectrode array,
Nanoparticles,
hiPSC,
3D engineered neural tissue,
Neurotoxicology
Conference:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays, Reutlingen, Germany, 4 Jul - 6 Jul, 2018.
Presentation Type:
Poster Presentation
Topic:
Stem cell-derived applications
Citation:
Mor
FM,
Roux
A,
Nikles
L,
Heuschkel
M and
Stoppini
L
(2019). Development of an in vitro model of NanoNeurotoxicity using human 3D neural tissues: Functional assay using porous microelectrode arrays.
Conference Abstract:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays.
doi: 10.3389/conf.fncel.2018.38.00052
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Received:
16 Mar 2018;
Published Online:
17 Jan 2019.
*
Correspondence:
Dr. Flavio M Mor, High School of Landscape Engineering and Architecture, University of Applied Sciences Western Switzerland, Geneva, 1202, Switzerland, flavio.mor@hesge.ch