Multiscale Organization of Neural Networks in a 3D Bioprinted Matrix

Area of Science:

• Neuroscience
• Biotechnology
• Biomaterials Science

Background:

• Accurate in vitro neural models are crucial for neuroscience research.
• 3D bioprinting offers high design freedom for complex neural constructs.
• Mature neurons require careful handling due to their delicate nature.

Purpose of the Study:

• To develop a 3D bioprinted neural model using gelatin methacryloyl (GelMA) and embryonic rat cortical neurons.
• To assess the structural and functional connectivity of the developed 3D neural network.
• To validate the model's utility in recapitulating in vivo conditions, including disease modeling.

Main Methods:

• Extrusion-based 3D bioprinting of GelMA-based constructs with E18 rat cortical neurons (3D neuMatrix).
• Assessment of neuronal viability and network formation.
• Bulk transcriptomic profiling to compare 3D neuMatrix with E18 cortex and 2D cultures.
• Establishment of an in vitro ischemic stroke model using the 3D neuMatrix.

Main Results:

• 3D neuMatrix demonstrated favorable neuronal viability and progressive formation of a 3D brain-like neural network.
• The model exhibited local and long-range functional axon connections.
• Transcriptomic analysis showed 3D neuMatrix closely resembles the E18 cortex.
• The 3D neuMatrix successfully modeled ischemic stroke, recapitulating viability, function, and transcriptomic features of MCAO/R.

Conclusions:

• 3D neuMatrix enables the formation of multiscale neural circuits in vitro.
• This model holds significant potential for studying neurodevelopment and neurological diseases.
• The 3D neuMatrix is a valuable platform for in vitro disease modeling, drug screening, and advancing in vitro intelligence.