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Stiff-to-Soft Transition from Glass to 3D Hydrogel Substrates in Neuronal Cell Culture
1Neuro-Nanoscale Engineering, Department of Mechanical Engineering and Institute of Complex Molecular Systems (ICMS), Eindhoven University of Technology, 5600 MB Eindhoven, The Netherlands.
Micromachines
|February 11, 2021
Summary
Researchers developed a 3D hydrogel brain model using gelatin methacryloyl (GelMA) to study neuronal behavior and mechanotransduction. The optimal cell seeding density was found to be between 1500-5000 cells/cm², enabling advanced brain-on-a-chip applications.
Area of Science:
- Biomaterials Science
- Neuroscience
- Tissue Engineering
Background:
- Hydrogels offer promising platforms for mimicking 3D brain architectures in vitro.
- Understanding in vitro human brain models and mechanotransduction is crucial for neurological research.
Purpose of the Study:
- To generate and characterize a 3D hydrogel model using GelMA and PEGDA for studying SH-SY5Y neuroblastoma cells.
- To investigate the effects of hydrogel properties and cell seeding density on neuronal morphology and migration.
- To establish an optimal cell seeding density for developing advanced 3D brain models.
Main Methods:
- Photo-polymerized GelMA and PEGDA hydrogels were cast atop SH-SY5Y neuroblastoma cells within PDMS rings.
- Cell morphology, survival, and migration were assessed using confocal z-stack microscopy.
- A systematic study of cell seeding densities was conducted to determine optimal ranges.
Main Results:
- GelMA hydrogels supported SH-SY5Y cell survival and promoted a neuron-like morphology with branching and spreading.
- PEGDA hydrogels did not support cell survival.
- Stiff-to-soft material transitions facilitated neuronal migration into the third dimension.
- Cell seeding densities above 10,000 cells/cm² led to aggregate formation, while densities below 1500 cells/cm² resulted in single cells.
Conclusions:
- GelMA hydrogels are suitable for creating 3D neuronal models.
- An optimal cell seeding density for GelMA hydrogels is between 1500 and 5000 cells/cm².
- This hydrogel construct advances the development of 3D mechanotransduction models for brain-on-a-chip applications.
