Molecular and Functional Characterization of Different BrainSphere Models for Use in Neurotoxicity Testing on Microelectrode Arrays

Author:

Hartmann Julia1ORCID,Henschel Noah1,Bartmann Kristina12,Dönmez Arif12,Brockerhoff Gabriele1,Koch Katharina12,Fritsche Ellen123ORCID

Affiliation:

1. IUF—Leibniz-Research Institute for Environmental Medicine, Auf’m Hennekamp 50, 40225 Duesseldorf, Germany

2. DNTOX GmbH, Gurlittstraße 53, 40223 Düsseldorf, Germany

3. Medical Faculty, Heinrich-Heine-University, Universitätsstraße 1, 40225 Düsseldorf, Germany

Abstract

The currently accepted methods for neurotoxicity (NT) testing rely on animal studies. However, high costs and low testing throughput hinder their application for large numbers of chemicals. To overcome these limitations, in vitro methods are currently being developed based on human-induced pluripotent stem cells (hiPSC) that allow higher testing throughput at lower costs. We applied six different protocols to generate 3D BrainSphere models for acute NT evaluation. These include three different media for 2D neural induction and two media for subsequent 3D differentiation resulting in self-organized, organotypic neuron/astrocyte microtissues. All induction protocols yielded nearly 100% NESTIN-positive hiPSC-derived neural progenitor cells (hiNPCs), though with different gene expression profiles concerning regional patterning. Moreover, gene expression and immunocytochemistry analyses revealed that the choice of media determines neural differentiation patterns. On the functional level, BrainSpheres exhibited different levels of electrical activity on microelectrode arrays (MEA). Spike sorting allowed BrainSphere functional characterization with the mixed cultures consisting of GABAergic, glutamatergic, dopaminergic, serotonergic, and cholinergic neurons. A test method for acute NT testing, the human multi-neurotransmitter receptor (hMNR) assay, was proposed to apply such MEA-based spike sorting. These models are promising tools not only in toxicology but also for drug development and disease modeling.

Funder

Ministry for innovation, science and research of the State of North-Rhine Westphalia, Germany

Danish Environmental Protection Agency

Horizon Europe project PARC

Publisher

MDPI AG

Subject

General Medicine

Reference148 articles.

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