HydroMEA: A 3D Hydrogel Based Microfluidic Device to Study Electrophysiology for Myelinated Nerve-on-Chip

Clement, B. F.; Pfister, C.; Kurer, T.; Labouesse, C.; Deshmukh, D. V.; Lehmann, J.; Paganella, L. G.; Ruff, T.; Dranseika, V.; Weaver, S.; Sommer, L.; Tibbitt, M. W.; Voros, J.; Tringides, C. M.

doi:10.1101/2025.07.24.666400

bioRxiv · 10.1101/2025.07.24.666400

HydroMEA: A 3D Hydrogel Based Microfluidic Device to Study Electrophysiology for Myelinated Nerve-on-Chip

Clement, B. F.·Pfister, C.·Kurer, T.·Labouesse, C.·Deshmukh, D. V.·Lehmann, J.·Paganella, L. G.·Ruff, T.·Dranseika, V.·Weaver, S.·Sommer, L.·Tibbitt, M. W.·Voros, J.·Tringides, C. M.

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Abstract

Engineered in vitro platforms are powerful systems to study information flow in the nervous system. While existing polydimethylsiloxane (PDMS)-based microfluidic platforms offer precise architectures, the cultured neurons grow on two-dimensional (2D) planar multielectrode arrays (MEA). To mimic the native microenvironment, where neurons grow in three-dimensional (3D) extracellular matrices (ECM), 3D hydrogels can be designed to encapsulate cells and enable physiologically-mimicked behaviors. Here, we describe hydroMEA, a 3D platform fabricated by placing PDMS microstructures on a high-density MEA and filled with a desired hydrogel, to offer controlled topologies, physiologically-relevant microenvironments, and real-time electrophysiological measurements. First, we developed a gelatin methacryloyl (GelMA) hydrogel with incorporated ECM components and tuned the mechanical properties to match those of nerve tissue. The hydrogel was able to support: 1) the growth of iPSC-derived sensory neurons (hSNs) for >100 days; 2) co-cultures of hSN with human embryonic stem cell-derived Schwann cells (hSCs), to enable reliable 3D myelination. Next, hydroMEA were prepared for topologically- defined 3D growth and myelination in designated compartments. Finally, electrophysiological evaluation of hSN-hSCs co-cultures revealed increased conduction speeds indicating functional myelin. This platform is a promising tool to study cell-cell interactions and to functionally evaluate the effect of pharmacological compounds in a more translational manner.

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Clement, B. F., Pfister, C., Kurer, T., Labouesse, C., Deshmukh, D. V., Lehmann, J., Paganella, L. G., Ruff, T., Dranseika, V., Weaver, S., Sommer, L., Tibbitt, M. W., Voros, J., Tringides, C. M.. 2025-07-31. HydroMEA: A 3D Hydrogel Based Microfluidic Device to Study Electrophysiology for Myelinated Nerve-on-Chip. https://doi.org/10.1101/2025.07.24.666400

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