Abstract
Background & Purpose
The development of human iPSC-derived neurons has significantly improved the predictive accuracy of preclinical neurotoxicity assays. While 2D culture models lack the complexity of real nerve tissue, engineered 3D organoids offer a more biomimetic solution for drug screening. To enhance preclinical testing, 28bio developed a 3D Human Nerve-on-a-Chip system that models peripheral nerve anatomy, enabling the evaluation of clinically relevant metrics such as nerve conduction velocity (NCV) and myelination. This model bridges the gap between traditional in vitro methods and in vivo studies by providing a more predictive platform for neurotoxicity testing.
Methods
- 3D Nerve-on-a-Chip Fabrication:
- Human iPSC-derived motor neuron spheroids were co-cultured with primary human Schwann cells in a dual hydrogel system to support robust axon growth.
- Schwann cells played a key role in the spheroid fabrication process, facilitating neuron growth and enhancing myelination.
- Morphological & Structural Analysis:
- Immunostaining confirmed the presence of key markers:
- S100 for Schwann cells
- βIII-tubulin for neurons
- DAPI for nuclear visualization
- Electrophysiological Testing:
- Nerve conduction velocity (NCV) was successfully measured in both mono-culture (neurons only) and co-culture (neurons + Schwann cells) spheroids.
- Structural analysis at multiple scales confirmed successful axon growth and Schwann cell myelination.
Results
- Schwann Cell Migration & Myelination:
- Schwann cells effectively migrated and supported axon myelination in the 3D system.
- Functional Assessment:
- The system successfully recorded NCV measurements, a gold-standard metric for peripheral nerve function.
- Enhanced Viability:
- The engineered nerve-on-a-chip maintained cell viability and structural integrity during extended culture periods.
Conclusion
The 3D Human Nerve-on-a-Chip platform effectively models human peripheral nerve anatomy and function, providing a clinically relevant alternative to in vivo testing. This innovative system enables:
- Accurate neurotoxicity screening
- Improved drug candidate evaluation
- Enhanced understanding of Schwann cell-driven myelination
Future work will focus on refining the model’s sensitivity for detecting subtle toxic effects and enhancing electrophysiology testing capabilities.
