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Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model
Published on: October 18, 2015
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Rapid generation of functional engineered 3D human neuronal assemblies: network dynamics evaluated by
L Muzzi1, D Di Lisa1, P Arnaldi1
1Department of Informatics, Bioengineering, Robotics, and Systems Engineering (DIBRIS), University of Genova, Genova, Italy.
Journal of Neural Engineering
|November 29, 2021
Summary
Researchers developed a novel 3D human neuronal network model using chitosan scaffolds and human-induced pluripotent stem cell-derived neurons. This engineered network enables controlled electrophysiological investigations, advancing biomedical applications.
Area of Science:
- Neuroscience
- Biomaterials Engineering
- Stem Cell Biology
Background:
- Developing functional 3D neuronal networks is crucial for advanced electrophysiological studies.
- Existing models often face limitations in structural control, cell composition, and long-term viability.
- Human-induced pluripotent stem cells (hiPSCs) offer a promising source for patient-specific neuronal models.
Purpose of the Study:
- To adapt a protocol for rapid generation of human neurons for 3D network construction.
- To create structurally controlled and compositionally defined 3D neuronal networks for electrophysiology.
- To evaluate the functional and structural characteristics of these novel 3D networks.
Main Methods:
- Utilized biocompatible chitosan microbeads as a scaffold for 3D network formation.
- Co-cultured excitatory neurons derived from hiPSCs with astrocytes.
- Employed a modified NgN2 differentiation protocol for controlled cell density and composition.
- Integrated 3D networks with 60-channel microelectrode arrays (MEAs) for electrophysiological monitoring.
- Compared 3D cultures with parallel 2D cultures on chitosan scaffolds.
Main Results:
- Sustained healthy and functional 3D cultures up to 60 days in vitro (DIV).
- Demonstrated hiPSC-derived neurons adhered to chitosan, forming stable 3D assemblies.
- Observed spontaneous neuronal network activity within two weeks of culture.
- 3D networks exhibited functional electrophysiological properties comparable to or exceeding 2D models.
Conclusions:
- Successfully developed a novel method for generating 3D engineered human neuronal cultures.
- This approach overcomes limitations of traditional 2D and 3D neuronal models.
- The 3D human neuronal networks coupled with MEAs hold significant potential for biomedical applications and drug discovery.
