A human integrated in vitro Neurotoxicity Safety Platform using hiPS neurons (Peri.4U neurons)
-
1
Natural and Medical Sciences Institute at the University Tübingen, Electrophysiology, Germany
-
2
Natural and Medical Sciences Institute at the University Tübingen, Electrophysiology, Germany
-
3
Axiogenesis, Germany
Motivation
In vitro pharmacology profiling of new chemical entities during early phases of drug discovery has recently become an essential tool to predict clinical adverse effects. While for cardiac safety testing high technology platforms are available, specific in vitro neurotoxic panels are not, or in-vivo models are used. However, correlations between animal and human data might be weak in some cases and animal studies are quite expensive, ethically questionable and require large amount of compounds.
We developed a platform based on the usage of human stem-cell derived neurons (Peri.4U neurons from Axiogenesis) to be able to directly address human targets. Compound related effects on the electrical activity of the Peri.4U neurons were assessed by performing field potential recordings using multiwell microelectrode array (MEA)- technology (Multi Channel Systems, Reutlingen).
Material and Methods
Previously frozen stem-cell derived neurons from Axiogenesis (Peri4U) were seeded on PEI and laminin coated coverslips (patch-clamp) and microelectrode array (MEA) chips, respectively at a concentration of 500k cells/cm2. Recordings were obtained between 8-20 days post thawing.
Results
Patch clamp recordings revealed the presence of three different types of cells, i.e. phasic (42%), tonic (27%) and intermediate firing (31%) neurons. Tonically firing cells could generate frequencies > 25 Hz following a 100 pA depolarizing current injection (1s). Action potentials could be blocked by TTX in all three neuronal types. Spontaneous activity was present in 11.2% of all neurons investigated, but in 43.5% of Type C neurons (tonic type).
When cultivated on the MEA chips, spontaneous activity of the Peri.4U neurons was recorded in 70% of the wells. The activity was burst-like in 77% of all recordings indicating the presence and establishment of a functional neuronal network. The activity of different electrophysiological parameters was highly variable between the different individual experiments, strengthening the need of data normalization to the respective control condition of each individual experiment.
When challenged with neurotoxic compounds, the results could be strongly linked to published data with a very high sensitivity.
Discussion and Conclusion
The MEA platform is a suitable tool for the investigation of the electrical activity of stem-cell derived neurons. Reference compounds with known neurotoxic potential were analyzed for their effect on neuronal network behavior, thereby demonstrating the potency of this in vitro Neurotoxicity Safety Platform to reliably detect and quantify neurotoxic compound actions.
Acknowledgements
The research leading to these results has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement n° 115439, resources of which are composed of financial contribution from the European Union's Seventh Framework Programme (FP7/2007-2013) and EFPIA companies’ in kind contribution. This publication reflects only the author’s views and neither the IMI JU nor EFPIA nor the European Commission are liable for any use that may be made of the information contained therein.
Support was also received from the BMBF-program KMUinnovativ: BiotechnologieBioChance, 0316162A, #0316162B
Keywords:
Pharmacology,
Neuronal activity,
stem cell derived neurons,
compound test,
field action potential,
neurotox
Conference:
MEA Meeting 2016 |
10th International Meeting on Substrate-Integrated Electrode Arrays, Reutlingen, Germany, 28 Jun - 1 Jul, 2016.
Presentation Type:
Poster Presentation
Topic:
MEA Meeting 2016
Citation:
Hess
D,
Kraushaar
U,
Cesare
P,
Rascher-Eggstein
G and
Guenther
E
(2016). A human integrated in vitro Neurotoxicity Safety Platform using hiPS neurons (Peri.4U neurons).
Front. Neurosci.
Conference Abstract:
MEA Meeting 2016 |
10th International Meeting on Substrate-Integrated Electrode Arrays.
doi: 10.3389/conf.fnins.2016.93.00046
Copyright:
The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers.
They are made available through the Frontiers publishing platform as a service to conference organizers and presenters.
The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated.
Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed.
For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions.
Received:
22 Jun 2016;
Published Online:
24 Jun 2016.
*
Correspondence:
Dr. Udo Kraushaar, Natural and Medical Sciences Institute at the University Tübingen, Electrophysiology, Reutlingen, Germany, udo.kraushaar@nmi.de