Development of a microelectrode array (MEA) based neurotoxicity assay for detecting the seizurogenic activity of novel drug candidates
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1
Hochschule Albstadt-Sigmaringen, Life Sciences Faculty, Germany
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2
NMI Technologie Transfer GmbH, Germany
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3
Natural and Medical Sciences Institute, Germany
During the early phase of drug development, the identification of possible side effects of novel drug candidates on the neuronal activity is very important. Until now most of the neurotoxicity assays are based on in vivo or in vitro animal models, which are problematic in terms of ethical issues and predictivity for humans. Over the last few years the continuous improvement of human iPSC derived neurons has increased their importance in this field, since they offer a great opportunity for the investigation of compound effects directly on a complex in vitro system of human origin.
By combining these cells with the microelectrode array (MEA) technique we can investigate the functional neurotoxicity of novel drug candidates on a neuronal network and thereby detect their seizurogenic activity.
In order to achieve this goal, we examined the effects of different coating and plating conditions on the development of the electrical activity of the human iPSC derived neurons (GlutaNeurons, Cellular Dynamics International CDI, US) that were cultured on 24-well glass Multiwell-MEAs (Multi Channel Systems MCS, GER). We tested three different coatings: 0.1% PEI, 0.07% PEI, which was diluted in a ready-to-use borate buffer, and the 0.07% PEI coating in combination with a pre-dotting with Laminin (10 µg/ml). The cells were dotted on the electrode fields of the MEAs in medium with a Laminin concentration of 10 µg/ml. The results showed no major differences between the tested coatings. Therefore, and because the handling of the 0.07% PEI coating is easier compared to the other tested coatings, we continued the study with the 0.07% PEI coating and explored the effects of different plating conditions. We examined the influences of a lower and a higher Laminin concentration in the dotting medium (10 µg/ml and 100 µg/ml) and in the medium (1 µg/ml and 33.3 µg/ml) that was used to fill-up the wells after the cells attached to the bottom of the wells.
The cells that were plated in the higher Laminin concentration showed an earlier outgrowth of neurites as well as an earlier increase of the electrical activity and the occurrence of synchronous bursting compared to the cells plated in the lower Laminin concentration.
The results suggest that there is a difference in the development of the cells in the tested plating conditions. The higher Laminin concentration led to an earlier outgrowth of neurites which could have favored an earlier formation of a neuronal network and thereby an earlier occurrence of synchronous burst activity. These findings have to be further investigated to establish a reliable neurotoxicity assay for detecting the seizurogenic activity of novel drug candidates in the future.
Acknowledgements
The authors would like to thank Dr. Sabine Lange (CDI) for providing the cells and useful information as well as Multi Channel Systems for providing the Glass Multiwell MEAs. This work was partially funded by the Baden-Württemberg Ministry of Science, Research and Art.
Keywords:
Assay development,
human iPSC derived neurons,
microelectrode array,
seizurogenic compounds,
Neurotoxicity
Conference:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays, Reutlingen, Germany, 4 Jul - 6 Jul, 2018.
Presentation Type:
Oral Presentation
Topic:
Assay development
Citation:
Loser
D,
Danker
T,
Möller
C,
Niedworok
A and
Kraushaar
U
(2019). Development of a microelectrode array (MEA) based neurotoxicity assay for detecting the seizurogenic activity of novel drug candidates.
Conference Abstract:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays.
doi: 10.3389/conf.fncel.2018.38.00016
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Received:
27 Mar 2018;
Published Online:
17 Jan 2019.
*
Correspondence:
Mr. Dominik Loser, Hochschule Albstadt-Sigmaringen, Life Sciences Faculty, Albstadt, Germany, dominik.loser@nmi.de