The effects of electroporation buffer composition on cell viability and electro-transfection efficiency

Area of Science:

• Biophysics
• Cell Biology
• Molecular Biology

Background:

• Electroporation uses electric fields to permeabilize cell membranes for delivering molecules.
• Cell viability and transfection efficiency depend heavily on experimental parameters, including buffer properties.

Purpose of the Study:

• To investigate how electroporation buffer composition affects cell viability and electro-transfection efficiency (eTE).
• To explore the role of specific ions and osmolality modifiers in optimizing electroporation protocols.

Main Methods:

• Systematic exploration of HEPES-based buffers with varying salts and sugars for 3T3 fibroblast electroporation.
• Application of electrical pulses maintaining constant energy or charge flux.
• Assessment of cell viability and eTE for plasmid DNA delivery.

Main Results:

• Applied pulse energy primarily determined cell viability.
• Magnesium (Mg2+)-based buffers significantly improved cell viability, potentially via ATPase activation.
• Lidocaine, an ATPase inhibitor, abolished the viability enhancement by Mg2+.
• Mg2+ buffers reduced eTE compared to potassium (K+)-based buffers.

Conclusions:

• Buffer composition and pulsing conditions are crucial for maximizing cell viability and eTE in electroporation.
• Mg2+ shows promise for enhancing cell survival during electroporation but requires careful optimization for efficient transfection.