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Bubble Perfusion Brain Slice Culture with Single-Droplet Stimulus Delivery in a 3D Printed Microfluidic Device
Genoveve G Gutierrez1, Richard J Ortiz2, Victoria Norman3
1Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico 88003, United States.
ACS Measurement Science Au
|October 20, 2025
Summary
This study introduces a 3D printed microfluidic device using bubble perfusion for ex vivo brain slice culture. The system maintains tissue viability and enables pharmacological studies, demonstrating its utility for receptor-binding ligand research.
Area of Science:
- Neuroscience
- Bioengineering
- Microfluidics
Background:
- Ex vivo tissue culture models physiological conditions, particularly for complex brain mechanisms.
- 3D printing accelerates microfluidic device development for ex vivo tissue culture.
- Miniaturization in ex vivo culture is limited by tissue explant dimensions.
Purpose of the Study:
- To develop a 3D printed microfluidic perfusion device for ex vivo brain slices.
- To utilize a novel bubble perfusion technique for controlled media delivery.
- To assess the viability and functionality of brain tissue in the system for pharmacological studies.
Main Methods:
- Designed and fabricated a 3D printed microfluidic device incorporating bubble perfusion.
- Integrated heating and prewarming chambers to maintain precise media droplet temperatures.
- Evaluated tissue viability using intracellular Ca2+ flux imaging and propidium iodide staining.
- Conducted pharmacological studies using cannabidiol (CBD) and anandamide (AEA) stimuli.
Main Results:
- Bubble perfusion delivered media droplets at a stable temperature (36.8 ± 0.13 °C) with minimal drift.
- Murine brain slices (SCN and EC) exhibited robust Ca2+ flux responses to KCl stimulus.
- Tissue remained viable for up to 12 hours, with approximately 60% of cells showing no membrane damage.
- CBD and AEA stimuli induced similar Ca2+ flux magnitudes and temporal dynamics.
Conclusions:
- The 3D printed bubble perfusion system supports ex vivo brain slice viability and function.
- The device is suitable for studying temporal dynamics of cellular responses to stimuli.
- This perfusion system is effective for pharmacological investigations of receptor-binding ligands.
