Intracellular mRNA Delivery via Lambda DNA-Based Viscoelastic Mechanoporation in Hyperbolic Microfluidic Channel

Area of Science:

• Biotechnology
• Cell Biology
• Microfluidics

Background:

• Intracellular delivery into suspension cells, especially immune cells like T- and B-lymphocytes, is a significant challenge.
• Existing microfluidic methods often require high-viscosity buffers, impacting cell viability and scalability.

Purpose of the Study:

• To develop a carrier-free microfluidic platform for efficient intracellular delivery into suspension cells.
• To overcome the limitations of high-viscosity buffers in current membrane disruption-based methods.

Main Methods:

• Utilized a hyperbolic microfluidic channel and a low-viscosity λDNA buffer for viscoelastic mechanoporation.
• Employed extensional strain to transiently deform cell membranes for cytosolic uptake.
• Investigated delivery mechanisms using Laurdan spectral analysis, ice incubation, and metabolic profiling.

Main Results:

• Achieved up to a 17-fold enhancement in mRNA delivery.
• Maintained over 85% cell viability across various suspension cell lines.
• Demonstrated tunable delivery efficiency through osmotic and cytoskeletal perturbations.

Conclusions:

• The viscoelastic mechanoporation platform enables efficient and safe mRNA and small molecule delivery into fragile immune cells.
• Understanding membrane dynamics is crucial for optimizing intracellular delivery outcomes.
• This tunable strategy offers a promising approach for advanced cell-based therapies.