Reversibly-sealable microfluidic platform for multi-molecule gradient delivery to large adherent cell cultures

Area of Science:

• Microfluidics
• Cell Biology
• Biomedical Engineering

Background:

• Spatial control of chemical gradients and flow is crucial for studying adherent cell behavior and drug responses.
• Existing microfluidic devices often have limitations like irreversible sealing and narrow shear stress ranges, restricting cell access and physiological relevance.

Purpose of the Study:

• To present a novel, reversibly sealable microfluidic platform for controlled delivery of small molecules to mammalian cells.
• To enable spatiotemporal control over chemical gradients and shear stress conditions across large cell culture areas.
• To provide a versatile and reusable tool for live-cell imaging and studying cellular responses.

Main Methods:

• Development of a reversibly sealable microfluidic device with a mechanical sandwich clamp for leak-free perfusion.
• Generation of tunable chemical gradients and a wide range of shear stresses (Stokes and laminar flow regimes) across centimeter-scale areas.
• Experimental and numerical analysis of molecule mixing, fluorophore uptake, and cell viability.

Main Results:

• The platform successfully generates robust, predictable chemical gradients and supports a wide range of shear stresses.
• Demonstrated ability to modulate mixing between co-flowing streams of small molecules.
• Verified fluorophore uptake and cell viability after perfusion and device removal.

Conclusions:

• The developed microfluidic platform offers a versatile and reusable solution for studying cellular responses to microenvironmental gradients.
• It overcomes limitations of existing devices by allowing cell access post-treatment and accommodating physiologically relevant shear stress conditions.
• This technology facilitates advanced research in cell biology and therapeutic response investigation.