Chemically induced synaptic activity between mixed primary hippocampal co-cultures in a microfluidic system
Graham Robertson1, Trevor J Bushell, Michele Zagnoni
1Department of Biomedical Engineering, University of Strathclyde, Glasgow, G4 0NW, UK.
Summary
This study developed a microfluidic device for connected yet isolated neuronal cultures. The system enables monitoring synaptic activity and investigating central nervous system disease models.
Area of Science:
- Neuroscience
- Biotechnology
Background:
- Primary neuronal cultures are crucial for studying central nervous system diseases.
- Existing methods lack the ability to precisely control and monitor communication between distinct neuronal populations.
Purpose of the Study:
- To develop and validate a novel microfluidic system for creating synaptically connected, environmentally isolated twin neuronal cultures.
- To establish a platform for investigating functional synaptic connectivity and activity modulation in vitro.
Main Methods:
- Utilized microfluidic devices to culture paired primary hippocampal neuronal/glia cells.
- Employed immunocytochemical staining (β-III-Tubulin, synaptophysin) to confirm neuronal connection and synapse formation.
- Applied calcium imaging and controlled chemical stimulation (KCl, glutamate) to monitor neuronal activity.
- Validated a numerical model to ensure no cross-contamination of chemical stimuli.
Main Results:
- Demonstrated successful co-culture of synaptically connected, isolated neuronal populations.
- Confirmed functional synaptic connectivity through calcium imaging and response to chemical stimuli.
- Observed significant increases in calcium events in connected cultures upon stimulation of one population.
Conclusions:
- The developed microfluidic system offers a novel methodology for probing functional synaptic connectivity in primary hippocampal co-cultures.
- This platform facilitates high-throughput investigation of synaptic activity modulation for drug discovery and disease modeling.
