Biomodulatory Effects of Molecular Delivery in Human T Cells Using 3D-Printed Acoustofluidic Devices

Area of Science:

• Biotechnology
• Cell Biology
• Bioengineering

Background:

• Cell-based therapies show promise for treating diseases like cancer.
• Current manufacturing relies on viral transduction, facing efficiency and time limitations.
• Non-viral transfection methods also struggle with variability and low delivery efficiency.

Purpose of the Study:

• To assess biological parameters influencing ultrasound-mediated molecular delivery to human T cells using acoustofluidics.
• To evaluate the potential of 3D-printed acoustofluidic devices for non-viral cell therapy processing.

Main Methods:

• Utilized flow cytometry and confocal imaging to analyze fluorescent molecule delivery (calcein, FITC-Dextran) to human T cells.
• Investigated effects of cell plating density, cell cycle phase, and calcium chelation (EGTA) on molecular delivery.
• Examined the role of the actin cytoskeleton and nuclear membrane permeability post-sonoporation.

Main Results:

• Lower cell plating densities (100,000 cells/mL) significantly improved molecular delivery.
• Cells in the S phase of the cell cycle exhibited enhanced intracellular delivery and viability.
• EGTA reduced molecular delivery, and actin cytoskeleton dynamics influenced membrane recovery.
• Acoustofluidic treatment demonstrated potential for nuclear membrane permeabilization.

Conclusions:

• 3D-printed acoustofluidic devices effectively enhance molecular delivery to human T cells.
• Optimized parameters like cell density and cell cycle phase improve non-viral transfection efficiency.
• This technology presents a promising avenue for advancing non-viral cell therapy manufacturing processes.