An in vitro platform for characterizing axonal electrophysiology of individual human iPSC-derived nociceptors
Neuropathic pain is characterized by aberrant activity of specific nociceptor populations, as demonstrated through functional assessments such as microneurography. Current treatments against severe forms of neuropathic pain demonstrate insufficient efficacy or lead to unwanted side effects as they f...
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| Veröffentlicht in: | Biosensors & bioelectronics Jg. 281; S. 117418 |
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| Sprache: | Englisch |
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England
Elsevier B.V
01.08.2025
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| ISSN: | 0956-5663, 1873-4235, 1873-4235 |
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| Abstract | Neuropathic pain is characterized by aberrant activity of specific nociceptor populations, as demonstrated through functional assessments such as microneurography. Current treatments against severe forms of neuropathic pain demonstrate insufficient efficacy or lead to unwanted side effects as they fail to specifically target the affected nociceptors. Tools that can recapitulate aspects of microneurography in vitro would enable a more targeted compound screening. Therefore, we developed an in vitro platform combining a CMOS-based high-density microelectrode array with a polydimethylsiloxane (PDMS) guiding microstructure that captures the electrophysiological responses of individual axons. Human induced pluripotent stem cell-derived (hiPSC) sensory neurons were cultured in a way that allowed axons to be distributed through parallel 4 ×10μm microchannels exiting the seeding well before converging to a bigger axon-collecting channel. This configuration allowed the measurement of stimulation-induced responses of individual axons. Sensory neurons were found to exhibit a great diversity of electrophysiological response profiles that can be classified into different functional archetypes. Moreover, we show that some responses are affected by applying the TRPV1 agonist capsaicin. Overall, results using our platform demonstrate that we were able to distinguish individual axon responses, making the platform a promising tool for testing therapeutic candidates targeting particular sensory neuron subtypes.
•Microfluidic platform on HD-MEA for recording hundreds of individually stimulated axons.•Axon conduction and excitability properties reveal distinct functional archetypes.•Distinct electrophysiological responses may correspond to neural subtypes. |
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| AbstractList | Neuropathic pain is characterized by aberrant activity of specific nociceptor populations, as demonstrated through functional assessments such as microneurography. Current treatments against severe forms of neuropathic pain demonstrate insufficient efficacy or lead to unwanted side effects as they fail to specifically target the affected nociceptors. Tools that can recapitulate aspects of microneurography in vitro would enable a more targeted compound screening. Therefore, we developed an in vitro platform combining a CMOS-based high-density microelectrode array with a polydimethylsiloxane (PDMS) guiding microstructure that captures the electrophysiological responses of individual axons. Human induced pluripotent stem cell-derived (hiPSC) sensory neurons were cultured in a way that allowed axons to be distributed through parallel 4 ×10μm microchannels exiting the seeding well before converging to a bigger axon-collecting channel. This configuration allowed the measurement of stimulation-induced responses of individual axons. Sensory neurons were found to exhibit a great diversity of electrophysiological response profiles that can be classified into different functional archetypes. Moreover, we show that some responses are affected by applying the TRPV1 agonist capsaicin. Overall, results using our platform demonstrate that we were able to distinguish individual axon responses, making the platform a promising tool for testing therapeutic candidates targeting particular sensory neuron subtypes. Neuropathic pain is characterized by aberrant activity of specific nociceptor populations, as demonstrated through functional assessments such as microneurography. Current treatments against severe forms of neuropathic pain demonstrate insufficient efficacy or lead to unwanted side effects as they fail to specifically target the affected nociceptors. Tools that can recapitulate aspects of microneurography in vitro would enable a more targeted compound screening. Therefore, we developed an in vitro platform combining a CMOS-based high-density microelectrode array with a polydimethylsiloxane (PDMS) guiding microstructure that captures the electrophysiological responses of individual axons. Human induced pluripotent stem cell-derived (hiPSC) sensory neurons were cultured in a way that allowed axons to be distributed through parallel 4 ×10μm microchannels exiting the seeding well before converging to a bigger axon-collecting channel. This configuration allowed the measurement of stimulation-induced responses of individual axons. Sensory neurons were found to exhibit a great diversity of electrophysiological response profiles that can be classified into different functional archetypes. Moreover, we show that some responses are affected by applying the TRPV1 agonist capsaicin. Overall, results using our platform demonstrate that we were able to distinguish individual axon responses, making the platform a promising tool for testing therapeutic candidates targeting particular sensory neuron subtypes.Neuropathic pain is characterized by aberrant activity of specific nociceptor populations, as demonstrated through functional assessments such as microneurography. Current treatments against severe forms of neuropathic pain demonstrate insufficient efficacy or lead to unwanted side effects as they fail to specifically target the affected nociceptors. Tools that can recapitulate aspects of microneurography in vitro would enable a more targeted compound screening. Therefore, we developed an in vitro platform combining a CMOS-based high-density microelectrode array with a polydimethylsiloxane (PDMS) guiding microstructure that captures the electrophysiological responses of individual axons. Human induced pluripotent stem cell-derived (hiPSC) sensory neurons were cultured in a way that allowed axons to be distributed through parallel 4 ×10μm microchannels exiting the seeding well before converging to a bigger axon-collecting channel. This configuration allowed the measurement of stimulation-induced responses of individual axons. Sensory neurons were found to exhibit a great diversity of electrophysiological response profiles that can be classified into different functional archetypes. Moreover, we show that some responses are affected by applying the TRPV1 agonist capsaicin. Overall, results using our platform demonstrate that we were able to distinguish individual axon responses, making the platform a promising tool for testing therapeutic candidates targeting particular sensory neuron subtypes. Neuropathic pain is characterized by aberrant activity of specific nociceptor populations, as demonstrated through functional assessments such as microneurography. Current treatments against severe forms of neuropathic pain demonstrate insufficient efficacy or lead to unwanted side effects as they fail to specifically target the affected nociceptors. Tools that can recapitulate aspects of microneurography in vitro would enable a more targeted compound screening. Therefore, we developed an in vitro platform combining a CMOS-based high-density microelectrode array with a polydimethylsiloxane (PDMS) guiding microstructure that captures the electrophysiological responses of individual axons. Human induced pluripotent stem cell-derived (hiPSC) sensory neurons were cultured in a way that allowed axons to be distributed through parallel 4 ×10μm microchannels exiting the seeding well before converging to a bigger axon-collecting channel. This configuration allowed the measurement of stimulation-induced responses of individual axons. Sensory neurons were found to exhibit a great diversity of electrophysiological response profiles that can be classified into different functional archetypes. Moreover, we show that some responses are affected by applying the TRPV1 agonist capsaicin. Overall, results using our platform demonstrate that we were able to distinguish individual axon responses, making the platform a promising tool for testing therapeutic candidates targeting particular sensory neuron subtypes. •Microfluidic platform on HD-MEA for recording hundreds of individually stimulated axons.•Axon conduction and excitability properties reveal distinct functional archetypes.•Distinct electrophysiological responses may correspond to neural subtypes. |
| ArticleNumber | 117418 |
| Author | Tringides, Christina M. Wallimann, Lea Duru, Jens Ruff, Tobias Petrella, Lorenzo Clément, Blandine F. Vörös, János |
| Author_xml | – sequence: 1 givenname: Blandine F. orcidid: 0000-0002-7305-0469 surname: Clément fullname: Clément, Blandine F. – sequence: 2 givenname: Lorenzo orcidid: 0009-0008-8416-995X surname: Petrella fullname: Petrella, Lorenzo – sequence: 3 givenname: Lea orcidid: 0009-0009-8233-8139 surname: Wallimann fullname: Wallimann, Lea – sequence: 4 givenname: Jens surname: Duru fullname: Duru, Jens – sequence: 5 givenname: Christina M. orcidid: 0000-0003-2747-1200 surname: Tringides fullname: Tringides, Christina M. – sequence: 6 givenname: János surname: Vörös fullname: Vörös, János email: janos.voros@biomed.ee.ethz.ch – sequence: 7 givenname: Tobias orcidid: 0000-0003-3565-5480 surname: Ruff fullname: Ruff, Tobias email: tobias.ruff@alumni.ethz.ch |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/40215890$$D View this record in MEDLINE/PubMed |
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| Keywords | Nociception Human iPSC-derived sensory neurons Axonal conduction CMOS-based microelectrode array In vitro model Activity-dependent slowing PDMS microstructures |
| Language | English |
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| SubjectTerms | Activity-dependent slowing agonists Axonal conduction axons Axons - drug effects Axons - physiology Biosensing Techniques - instrumentation biosensors capsaicin Capsaicin - pharmacology Cells, Cultured CMOS-based microelectrode array Electrophysiological Phenomena electrophysiology Equipment Design Human iPSC-derived sensory neurons Humans In vitro model Induced Pluripotent Stem Cells - cytology Microelectrodes microstructure Nociception Nociceptors - cytology Nociceptors - drug effects Nociceptors - physiology pain PDMS microstructures polydimethylsiloxane sensory neurons sowing therapeutics transient receptor potential vanilloid channels TRPV Cation Channels - agonists |
| Title | An in vitro platform for characterizing axonal electrophysiology of individual human iPSC-derived nociceptors |
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