Cell electrofusion based on nanosecond/microsecond pulsed electric fields.

Area of Science:

• Biophysics
• Cell Biology
• Electrical Engineering

Background:

• Microsecond pulsed electric fields are standard for cell electrofusion but struggle with cells of different sizes.
• Nanosecond pulses reduce size dependency but induce small, easily recoverable pores.
• Existing methods face challenges in achieving efficient fusion across diverse cell populations.

Purpose of the Study:

• To develop an innovative cell electrofusion method combining nanosecond and microsecond pulsed electric fields.
• To overcome the limitations of size-dependent electroporation in traditional methods.
• To enhance cell fusion efficiency by optimizing pore formation and distribution.

Main Methods:

• Finite element method simulations were employed to model pore distribution, radius, and density.
• A hybrid pulsed electric field protocol involving nanosecond and microsecond pulses was designed.
• A computational model of two contacting cells of different sizes was utilized.

Main Results:

• The nanosecond/microsecond pulse combination resulted in a large pore radius (70 nm) and high density (5×10^13 m^-2) specifically at the cell junction.
• Pores in non-contact areas remained small (1-10 nm) and sparse (10^9-10^12 m^-2).
• High transmembrane voltage (>1V) was localized to the cell junction, while other areas experienced lower voltage (≤0.6V).

Conclusions:

• The proposed nanosecond/microsecond pulsed electric field strategy significantly enhances cell fusion efficiency.
• Concentrating electroporation in the cell junction area is key to successful fusion of cells with varying sizes.
• This hybrid approach offers a promising solution for overcoming size-related challenges in cell electrofusion technology.