Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A...

Area of Science:

• Biotechnology
• Molecular Biology
• Oncology

Background:

• Electroporation uses electric fields for non-viral gene transfer, showing potential for cancer treatment.
• Gene electrotransfer is effective in vitro but faces challenges in complex 3D tumor environments.

Purpose of the Study:

• To compare gene electrotransfer efficiency using different pulsed electric field protocols in 2D and 3D cell cultures.
• To investigate the impact of pulse parameters on gene delivery in multicellular spheroids.

Main Methods:

• Comparison of electrochemotherapy and gene electrotherapy pulsed electric field protocols.
• Assessment of "High Voltage-Low Voltage" pulse variations.
• Evaluation of cell permeabilization and gene delivery in 2D cell suspensions and 3D multicellular spheroids.

Main Results:

• All tested protocols effectively permeabilized both 2D and 3D cells.
• Gene electrotransfer protocols showed higher efficiency in cell suspensions (approx. 50% transfection rate).
• Despite uniform permeabilization in 3D spheroids, gene delivery was limited to the spheroid periphery.

Conclusions:

• Electric field intensity and cell permeabilization are crucial for gene electrotransfer.
• Pulse duration is critical, as it affects plasmid electrophoretic mobility, which is hindered in 3D structures.
• Current gene electrotransfer methods struggle to deliver genes into the core of 3D tumor models.