Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arrays
The recent availability of human induced pluripotent stem cells (hiPSCs) holds great promise as a novel source of human-derived neurons for cell and tissue therapies as well as for in vitro drug screenings that might replace the use of animal models. However, there is still a considerable lack of kn...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2016-03-01
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| Series: | Frontiers in Neuroscience |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fnins.2016.00121/full |
| _version_ | 1828390553037832192 |
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| author | Hayder eAmin Alessandro eMaccione Federica eMarinaro Stefano eZordan Thierry eNieus Luca eBerdondini |
| author_facet | Hayder eAmin Alessandro eMaccione Federica eMarinaro Stefano eZordan Thierry eNieus Luca eBerdondini |
| author_sort | Hayder eAmin |
| collection | DOAJ |
| description | The recent availability of human induced pluripotent stem cells (hiPSCs) holds great promise as a novel source of human-derived neurons for cell and tissue therapies as well as for in vitro drug screenings that might replace the use of animal models. However, there is still a considerable lack of knowledge on the functional properties of hiPSC-derived neuronal networks, thus limiting their application. Here, upon optimization of cell culture protocols, we demonstrate that both spontaneous and evoked electrical spiking activities of these networks can be characterized on-chip by taking advantage of the resolution provided by CMOS multielectrode arrays (CMOS-MEAs). These devices feature a large and closely-spaced array of 4096 simultaneously recording electrodes and multi-site on-chip electrical stimulation. Our results show that networks of human-derived neurons can respond to electrical stimulation with a physiological repertoire of spike waveforms after three months of cell culture, a period of time during which the network undergoes the expression of developing patterns of spontaneous spiking activity. To achieve this, we have investigated the impact on the network formation and on the emerging network-wide functional properties induced by different biochemical substrates, i.e. poly-dl-ornithine (PDLO), poly-l-ornithine (PLO), and polyethylenimine (PEI), that were used as adhesion promoters for the cell culture. Interestingly, we found that neuronal networks grown on PDLO coated substrates show significantly higher spontaneous firing activity, reliable responses to low-frequency electrical stimuli, and an appropriate level of PSD-95 that may denote a physiological neuronal maturation profile and synapse stabilization. However, our results also suggest that even three-month culture might not be sufficient for human-derived neuronal network maturation. Taken together, our results highlight the tight relationship existing between substrate coatings and emerging network properties, i.e. spontaneous activity, responsiveness, synapse formation and maturation. Additionally, our results provide a baseline on the functional properties expressed over three months of network development for a commercially available line of hiPSC-derived neurons. This is a first step toward the development of functional pre-clinical assays to test pharmaceutical compounds on human-derived neuronal networks with CMOS-MEAs. |
| first_indexed | 2024-12-10T06:46:18Z |
| format | Article |
| id | doaj.art-7ccf9b11264d4272bc34a601b30ac0d6 |
| institution | Directory Open Access Journal |
| issn | 1662-453X |
| language | English |
| last_indexed | 2024-12-10T06:46:18Z |
| publishDate | 2016-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Neuroscience |
| spelling | doaj.art-7ccf9b11264d4272bc34a601b30ac0d62022-12-22T01:58:38ZengFrontiers Media S.A.Frontiers in Neuroscience1662-453X2016-03-011010.3389/fnins.2016.00121183111Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arraysHayder eAmin0Alessandro eMaccione1Federica eMarinaro2Stefano eZordan3Thierry eNieus4Luca eBerdondini5Fondazione Istituto Italiano di Tecnologia (IIT)Fondazione Istituto Italiano di Tecnologia (IIT)Fondazione Istituto Italiano di Tecnologia (IIT)Fondazione Istituto Italiano di Tecnologia (IIT)Fondazione Istituto Italiano di Tecnologia (IIT)Fondazione Istituto Italiano di Tecnologia (IIT)The recent availability of human induced pluripotent stem cells (hiPSCs) holds great promise as a novel source of human-derived neurons for cell and tissue therapies as well as for in vitro drug screenings that might replace the use of animal models. However, there is still a considerable lack of knowledge on the functional properties of hiPSC-derived neuronal networks, thus limiting their application. Here, upon optimization of cell culture protocols, we demonstrate that both spontaneous and evoked electrical spiking activities of these networks can be characterized on-chip by taking advantage of the resolution provided by CMOS multielectrode arrays (CMOS-MEAs). These devices feature a large and closely-spaced array of 4096 simultaneously recording electrodes and multi-site on-chip electrical stimulation. Our results show that networks of human-derived neurons can respond to electrical stimulation with a physiological repertoire of spike waveforms after three months of cell culture, a period of time during which the network undergoes the expression of developing patterns of spontaneous spiking activity. To achieve this, we have investigated the impact on the network formation and on the emerging network-wide functional properties induced by different biochemical substrates, i.e. poly-dl-ornithine (PDLO), poly-l-ornithine (PLO), and polyethylenimine (PEI), that were used as adhesion promoters for the cell culture. Interestingly, we found that neuronal networks grown on PDLO coated substrates show significantly higher spontaneous firing activity, reliable responses to low-frequency electrical stimuli, and an appropriate level of PSD-95 that may denote a physiological neuronal maturation profile and synapse stabilization. However, our results also suggest that even three-month culture might not be sufficient for human-derived neuronal network maturation. Taken together, our results highlight the tight relationship existing between substrate coatings and emerging network properties, i.e. spontaneous activity, responsiveness, synapse formation and maturation. Additionally, our results provide a baseline on the functional properties expressed over three months of network development for a commercially available line of hiPSC-derived neurons. This is a first step toward the development of functional pre-clinical assays to test pharmaceutical compounds on human-derived neuronal networks with CMOS-MEAs.http://journal.frontiersin.org/Journal/10.3389/fnins.2016.00121/fullneural networksSurface functionalizationIPSC-derived neuronsspontaneous and evoked activitiesCMOS-multielectrode arrays |
| spellingShingle | Hayder eAmin Alessandro eMaccione Federica eMarinaro Stefano eZordan Thierry eNieus Luca eBerdondini Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arrays Frontiers in Neuroscience neural networks Surface functionalization IPSC-derived neurons spontaneous and evoked activities CMOS-multielectrode arrays |
| title | Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arrays |
| title_full | Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arrays |
| title_fullStr | Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arrays |
| title_full_unstemmed | Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arrays |
| title_short | Electrical responses and spontaneous activity of human iPS-derived neuronal networks characterized for three-month culture with 4096-electrode arrays |
| title_sort | electrical responses and spontaneous activity of human ips derived neuronal networks characterized for three month culture with 4096 electrode arrays |
| topic | neural networks Surface functionalization IPSC-derived neurons spontaneous and evoked activities CMOS-multielectrode arrays |
| url | http://journal.frontiersin.org/Journal/10.3389/fnins.2016.00121/full |
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