Parallelization of MEA-based electrophysiological recordings of murine and human islets of Langerhans
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1
Natural and Medical Sciences Institute at the University Tübingen, Electrophysiology
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2
Multi Channel Systems, Germany
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3
Tübingen University, Institute of Pharmacy, Pharmacology, Germany
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4
Tübingen University, Institute of Pharmacy, Pharmacology, Germany
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5
Natural and Medical Sciences Institute at the University Tübingen, Electrophysiology, Germany
Motivation
The increase in postprandial blood glucose concentration leads to glucose-induced electrical activity of pancreatic beta-cells resulting in insulin secretion. This electrical activity typically manifests in glucose concentration-dependent oscillations which can easily be recorded by microelectrode arrays (MEA). Until now the number of experiments was limited to one at the time. To increase the throughput to an extent suitable for drug development we developed a MEA-based parallelized recording system for islets of Langerhans.
Material and Methods
Intact murine or human islets of Langerhans were used acutely on the 3D-Cluster Screen. Electrical oscillatory activity was repetitively recorded at 37°C as field potentials and quantified as fraction of plateau phase (FOPP = percentage of time with burst activity) for murine islets. In case of human islets, overall spike activity was used. Chips consisting of TiN electrodes on polyimide foils supported by a ceramic plate were developed in the clean room of the NMI, whereas the amplifier was a modified MEA2100 system from Multi Channel Systems.
Results
Glucose-dependent electrical oscillatory activity in beta-cells within islets of Langerhans is important for understanding their physiology and pathophysiology. Electrophysiological recordings are both time consuming and technically challenging. Until now electrophysiological investigations of islets of Langerhans were restricted to a single recording per experiment. In order to meet the requirements necessary for drug screening and profiling, we developed a MEA-based high-throughput system for the recording of acute intact islets of Langerhans, the 3D-Cluster Screen. The recording chip consists of five chambers which allow to record electrical oscillations from up to five intact islets of Langerhans simultaneously per chamber. Therefore the current layout of the 3D-Cluster Screen MEA chip allows to record electrical oscillations from up to 25 intact islets of Langerhans simultaneously. Positioning of the islets is realized by suction of the islets onto the electrodes. All five chambers can be perfused independently in order to perform up to five individual experiments in parallel. This allowes to achieve e.g. concentration response-curves within 2 h with more than 10 repeats.
Discussion and Conclusion
We increased the throughput from MEA-based acutely recorded islets of Langerhans from one single recording per experiment up to 25 recordings with the 3D-Cluster Screen device. It transfers this electrophysiological approach from the academic to the industrial level, allowing to integrate this method into drug screening and profiling.
Glucose-dependent electrical activity could be recorded from both primary murine and human islets of Langerhans for more than three hours. Validation experiments including glucose concentration-response curves and application of different ion channel modulators affirm that the physiological behavior is unaltered by this recording method. The development of the parallelized system opens the door for large-scale investigations of beta-cell electrical activity.
Due to its versatile usability the 3D-Cluster screen is not limited to primary islets of Langerhans but is also suitable for stem-cell derived tissue-like 3D structures like beta-cell and cardiac clusters.
igure legends
Figure 1: Layout of the 3D-Cluster Screen device. 3D-Cluster Screen MEA design with 5 x 5 recording electrodes. This layout allows simultaneous recordings from up to 25 intact islets of Langerhans. Multiwell cover with five recording chambers on top of the new 3D-Cluster Screen MEA chip. Each of the five chambers can be perfused independently which allows five independent experiments per recording.
Figure 2: Magnification of the electrode field. A-C: Each of the five electrodes is surrounded by several suction holes. Gentle negative pressure from below the chip positions individual islets of Langerhans on top of an electrode. D: in this well five out of five electrodes are occupied by individual islets of Langerhans. E: Close-up onto one islet sitting on top of the electrode. For visualization suction holes are marked as circles.
Figure 3: Exemplary recording. In this typical example data from up to five islets per well could be recorded in parallel from four out of five wells (well 1 was deactivated).
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:
Insulin,
Pancreas,
beta cells,
Oscillatory activity,
parallelization,
throughput
Conference:
MEA Meeting 2016 |
10th International Meeting on Substrate-Integrated Electrode Arrays, Reutlingen, Germany, 28 Jun - 1 Jul, 2016.
Presentation Type:
oral
Topic:
MEA Meeting 2016
Citation:
Kraushaar
U,
Schönecker
S,
Krippeit-Drews
P,
Drews
G and
Guenther
E
(2016). Parallelization of MEA-based electrophysiological recordings of murine and human islets of Langerhans.
Front. Neurosci.
Conference Abstract:
MEA Meeting 2016 |
10th International Meeting on Substrate-Integrated Electrode Arrays.
doi: 10.3389/conf.fnins.2016.93.00018
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Received:
22 Jun 2016;
Published Online:
24 Jun 2016.
*
Correspondence:
Dr. Udo Kraushaar, Natural and Medical Sciences Institute at the University Tübingen, Electrophysiology, Reutlingen, udo.kraushaar@nmi.de