Non-contact monitoring of extra-cellular field potentials with a multi-electrode array.

Developing tools to enable non-invasive, high-throughput electrophysiology measurements of large functional-networks of electrogenic cells used as in vitro disease models for the heart and brain remains an outstanding challenge for preclinical drug discovery, where failures are costly and can prove to be fatal during clinical trials. Here we demonstrate, for the first time, that it is possible to perform non-contact monitoring of extra-cellular field potentials with a multi-electrode array (MEA). To do this preliminary demonstration we built a prototype with a custom mechanical stage to micro-position cells grown on conventional glass coverslips over the recording surface of a MEA sensor. The prototype can monitor extra-cellular fields generated by multi-cellular networks in a non-contact configuration, enabling a single MEA sensor to probe different cultures in succession, without fouling or degrading its sensitive electronic surface. This first demonstration with easy to culture cardiomyocyte cells and a prototype device points to the exciting possibility for instrument development leading to more efficient and cost-effective drug screening paradigms for cardiovascular and neurological diseases.

[1]  Ofer Feinerman,et al.  Reliable neuronal logic devices from patterned hippocampal cultures , 2008 .

[2]  D. A. Dunnett.  Classical Electrodynamics , 2020, Nature.

[3]  Alon Spira,et al.  High-Resolution Electrophysiological Assessment of Human Embryonic Stem Cell-Derived Cardiomyocytes: A Novel In Vitro Model for the Study of Conduction , 2002, Circulation research.

[4]  G. Whitesides,et al.  Self-Assembled Monolayers That Resist the Adsorption of Proteins and the Adhesion of Bacterial and Mammalian Cells , 2001 .

[5]  Bruce C. Wheeler,et al.  Designing Neural Networks in Culture , 2010, Proceedings of the IEEE.

[6]  Paul S. Weiss,et al.  The Brain Activity Map , 2013, Science.

[7]  R. Passier,et al.  Expansion and patterning of cardiovascular progenitors derived from human pluripotent stem cells , 2015, Nature Biotechnology.

[8]  Benoit Charlot,et al.  An integrated microfluidic/microelectrode array for the study of activity-dependent intracellular dynamics in neuronal networks. , 2018, Lab on a chip.

[9]  Sean P Sheehy,et al.  Neurons derived from different brain regions are inherently different in vitro: a novel multiregional brain-on-a-chip. , 2017, Journal of neurophysiology.

[10]  R. Peri,et al.  High-throughput electrophysiology: an emerging paradigm for ion-channel screening and physiology , 2008, Nature Reviews Drug Discovery.

[11]  M. Baudry,et al.  Advances in Network Electrophysiology , 2006 .

[12]  D. Tampellini,et al.  Synaptic activity and Alzheimer's disease: a critical update , 2015, Front. Neurosci..

[13]  P. Doevendans,et al.  Developmental changes in ionic channel activity in the embryonic murine heart. , 1996, Circulation research.

[14]  Rafael Yuste,et al.  Nanotools for neuroscience and brain activity mapping. , 2013, ACS nano.

[15]  Rama S. Verma,et al.  An improved protocol for primary culture of cardiomyocyte from neonatal mice , 2008, In Vitro Cellular & Developmental Biology - Animal.

[16]  R C Barr,et al.  Electrophysiological effects of remodeling cardiac gap junctions and cell size: experimental and model studies of normal cardiac growth. , 2000, Circulation research.

[17]  L. Formigli,et al.  Skeletal myoblasts overexpressing relaxin improve differentiation and communication of primary murine cardiomyocyte cell cultures. , 2009, Journal of molecular and cellular cardiology.

[18]  Mohammad Amin Sherafat,et al.  Generation of serotonin neurons from human pluripotent stem cells , 2016, Nature Biotechnology.

[19]  Bian Wu,et al.  MEA Viewer: A high-performance interactive application for visualizing electrophysiological data , 2017, bioRxiv.

[20]  Rona S. Gertner,et al.  CMOS nanoelectrode array for all-electrical intracellular electrophysiological imaging. , 2017, Nature nanotechnology.

[21]  Sean P Sheehy,et al.  A linked organ-on-chip model of the human neurovascular unit reveals the metabolic coupling of endothelial and neuronal cells , 2018, Nature Biotechnology.

[22]  Qixing Huang,et al.  Use of RNAi technology to develop a PRSV-resistant transgenic papaya , 2017, Scientific Reports.

[23]  Jochen Guck,et al.  Mechanosensing is critical for axon growth in the developing brain , 2016, Nature Neuroscience.

[24]  Jerome Pine,et al.  Extracellular potentials in low-density dissociated neuronal cultures , 2002, Journal of Neuroscience Methods.

[25]  Deepak Srivastava,et al.  Contractility of single cardiomyocytes differentiated from pluripotent stem cells depends on physiological shape and substrate stiffness , 2015, Proceedings of the National Academy of Sciences.

[26]  U. Frey,et al.  Tracking axonal action potential propagation on a high-density microelectrode array across hundreds of sites , 2013, Nature Communications.

[27]  Bruce C Wheeler,et al.  Novel MEA platform with PDMS microtunnels enables the detection of action potential propagation from isolated axons in culture. , 2009, Lab on a chip.

[28]  Gordana Vunjak-Novakovic,et al.  Advanced maturation of human cardiac tissue grown from pluripotent stem cells , 2018, Nature.

[29]  Donald M Bers,et al.  Screening Drug-Induced Arrhythmia Using Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes and Low-Impedance Microelectrode Arrays , 2013, Circulation.

[30]  Peter Molnar,et al.  Patterned cardiomyocytes on microelectrode arrays as a functional, high information content drug screening platform. , 2011, Biomaterials.

[31]  C. Kahn,et al.  Regulation of Myocardial Contractility and Cell Size by Distinct PI3K-PTEN Signaling Pathways , 2002, Cell.

[32]  Ulrich Egert,et al.  Cellular Physiology Cellular Physiology Cellular Physiology Cellular Physiology Cellular Physiology Estimation of Action Potential Changes from Field Potential Recordings in Multicellular Mouse Cardiac Myocyte Cultures Key Words Microelectrode Array @bullet Action Potential Upstroke @bullet Action P , 2022 .

[33]  R. Passier,et al.  Interpretation of field potentials measured on a multi electrode array in pharmacological toxicity screening on primary and human pluripotent stem cell-derived cardiomyocytes , 2017, Biochemical and biophysical research communications.

[34]  C. Koch,et al.  On the origin of the extracellular action potential waveform: A modeling study. , 2006, Journal of neurophysiology.

[35]  Kenneth S Kosik,et al.  Diseases: Study neuron networks to tackle Alzheimer's , 2013, Nature.

[36]  D. Huh,et al.  Organs-on-chips at the frontiers of drug discovery , 2015, Nature Reviews Drug Discovery.

[37]  Kenneth L. Shepard,et al.  A very large-scale microelectrode array for cellular-resolution electrophysiology , 2017, Nature Communications.

[38]  Donald E Ingber,et al.  Organs-on-Chips with combined multi-electrode array and transepithelial electrical resistance measurement capabilities. , 2017, Lab on a chip.

[39]  D. Ingber,et al.  Microfluidic organs-on-chips , 2014, Nature Biotechnology.

[40]  C. Koch,et al.  The origin of extracellular fields and currents — EEG, ECoG, LFP and spikes , 2012, Nature Reviews Neuroscience.

[41]  Vijay Viswam,et al.  High-resolution CMOS MEA platform to study neurons at subcellular, cellular, and network levels. , 2015, Lab on a chip.

[42]  Ulrich Egert,et al.  Biological application of microelectrode arrays in drug discovery and basic research , 2003, Analytical and bioanalytical chemistry.

[43]  A K Capulli,et al.  Approaching the in vitro clinical trial: engineering organs on chips. , 2014, Lab on a chip.

[44]  R. Niwa,et al.  Low-frequency extracellular potentials recorded from the sinoatrial node. , 1998, Cardiovascular research.

[45]  Kristofer E. Bouchard,et al.  Use of a Neural Circuit Probe to Validate in silico Predictions of Inhibitory Connections , 2017, bioRxiv.

[46]  A. Offenhäusser,et al.  Electrical recordings from rat cardiac muscle cells using field-effect transistors. , 1999, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[47]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

[48]  Paul K. Hansma,et al.  Electromagnetic squeezer for compressing squeezable electron tunneling junctions. Technical report , 1984 .

[49]  Bartosz A. Grzybowski,et al.  Directing cell motions on micropatterned ratchets , 2009 .

[50]  G. Pazour,et al.  Ror2 signaling regulates Golgi structure and transport through IFT20 for tumor invasiveness , 2017, Scientific Reports.

[51]  F. J. Livesey,et al.  Guided self-organization and cortical plate formation in human brain organoids , 2017, Nature Biotechnology.

[52]  Jonathan A. Bernstein,et al.  Assembly of functionally integrated human forebrain spheroids , 2017, Nature.

[53]  M. Spira,et al.  Multi-electrode array technologies for neuroscience and cardiology. , 2013, Nature nanotechnology.

[54]  N. Ziv,et al.  Evolution of Action Potential Propagation and Repolarization in Cultured Neonatal Rat Ventricular Myocytes , 2001, Journal of cardiovascular electrophysiology.

[55]  M. Peschanski,et al.  Combinatorial analysis of developmental cues efficiently converts human pluripotent stem cells into multiple neuronal subtypes , 2014, Nature Biotechnology.

[56]  Udi Nussinovitch,et al.  Sinoatrial node cardiomyocytes derived from human pluripotent cells function as a biological pacemaker , 2016, Nature Biotechnology.

[57]  Heribert Bohlen,et al.  Determination of electrical properties of ES cell-derived cardiomyocytes using MEAs. , 2004, Journal of electrocardiology.

[58]  D. Dumitru,et al.  Physiologic basis of potentials recorded in electromyography , 2000, Muscle & nerve.