Modeling the Blood-Brain Barrier in a 3D triple co-culture microfluidic system.

Area of Science:

• Neuroscience
• Biomedical Engineering
• Cell Biology

Background:

• Accurate in-vitro blood-brain barrier (BBB) models are crucial for research.
• Existing models lack the ability to study neuronal responses to blood-borne factors crossing the BBB.
• There is a need for advanced models that mimic in-vivo physiological conditions.

Purpose of the Study:

• To establish a 3D triple co-culture microfluidic system that accurately replicates the in-vivo blood-brain barrier (BBB).
• To enable the study of neuronal growth and function influenced by substances crossing the BBB.
• To provide a more physiologically relevant platform for drug discovery and neurological research.

Main Methods:

• Development of a 3D microfluidic system utilizing human umbilical vein endothelial cells (HUVEC), primary rat astrocytes, and neurons.
• Immunostaining to confirm the integrity and tight intercellular junctions of the endothelial monolayer, establishing an intact BBB.
• Fluorescent-based assays with varying molecular weight dextrans to assess BBB selective permeability.
• Calcium imaging to demonstrate and validate neuron functionality within the BBB model.

Main Results:

• Successful establishment of a 3D triple co-culture microfluidic system.
• Confirmation of an intact blood-brain barrier (BBB) with tight intercellular junctions via immunostaining.
• Demonstration of selective permeability characteristic of the BBB using dextran assays.
• Validation of neuronal functionality through calcium imaging.

Conclusions:

• The developed 3D triple co-culture microfluidic system effectively mimics the in-vivo blood-brain barrier (BBB).
• This novel model allows for the investigation of neuronal responses to blood-borne factors, addressing a significant gap in current research.
• The system offers a promising platform for advancing neurological studies and therapeutic development.