Selective electrofusion of conjugated cells in flow

Area of Science:

• Biotechnology
• Cell Biology
• Biophysics

Background:

• Cell fusion is essential for various biological processes and therapeutic applications.
• Traditional cell fusion methods can be inefficient and lack precise control.
• Developing high-throughput, controlled cell fusion techniques is critical for advancing cell-based therapies.

Purpose of the Study:

• To develop and validate a novel flow cytometry-based method for inducing electrofusion of K562 and L1210 cells.
• To optimize electric pulse parameters for efficient cell fusion in a continuous flow system.
• To assess the efficacy of electrofusion using advanced microscopy techniques.

Main Methods:

• Utilized a modified flow cytometer integrating hydrodynamic focusing and an electric pulse application system.
• Employed fluorescent membrane probes (DiO and DiI) for optical recognition of K562 and L1210 cell pairs.
• Established an avidin-biotin bridge for specific cell coupling prior to electrofusion.
• Analyzed fused cells using normal and confocal microscopy to quantify fusion rates.

Main Results:

• Successfully induced electrofusion of K562 and L1210 cells within a flow cytometer.
• Achieved fusion in approximately 10% of pulsed cell pairs under optimized conditions.
• Determined optimal electric pulse parameters: 10-15 microseconds duration and 4-8 x 10^5 V/m field strength.
• Demonstrated the feasibility of high-throughput, optically monitored cell fusion.

Conclusions:

• The developed flow cytometry-based electrofusion method offers a controlled and efficient approach for cell fusion.
• This technique has potential applications in drug screening, cell-based therapies, and synthetic biology.
• Further optimization may enhance fusion efficiency and broaden the applicability of this method.