High throughput 3D gel-based neural organotypic model for cellular assays using fluorescence biosensors

Area of Science:

• Neuroscience
• Biotechnology
• Pharmacology

Background:

• Three-dimensional (3D) organotypic models are crucial for developing clinically predictive assays in therapeutics development.
• Existing models struggle to fully replicate native-like physiological features of tissues, especially for neurological diseases.
• Biofabrication of 3D gel-based systems offers a versatile approach for creating complex, native-like tissue models.

Purpose of the Study:

• To develop functional 3D neural co-culture models using biofabrication techniques.
• To integrate biosensors and optogenetics for real-time neurotransmitter and intracellular calcium measurements.
• To assess the predictive power of these 3D models for drug responses compared to 2D models.

Main Methods:

• Fabrication of 3D fibrin gel-based neural co-cultures with human induced pluripotent stem cell (hiPSC)-derived dopaminergic/glutamatergic neurons and astrocytes.
• Incorporation of genetically encoded fluorescence biosensors for monitoring intracellular calcium, dopamine, and glutamate.
• Application of optogenetics for neural activation and pharmacological perturbations for drug response assessment.

Main Results:

• Successful creation of functional 3D neural co-culture models with key cell types.
• Real-time monitoring of intracellular calcium and neurotransmitter dynamics achieved using biosensors and optogenetics.
• 3D models demonstrated drug responses consistent with in-vivo data, outperforming 2D models in certain cases.

Conclusions:

• 3D fibrin gel-based neural models offer a physiologically relevant platform for studying neurological diseases.
• These advanced models provide a more accurate preclinical assessment of drug efficacy and toxicity.
• The developed platform supports high-throughput screening and advances therapeutics development for neurological disorders.