Microfluidic\nChip for Cell Fusion and In Situ Separation\nof Fused Cells

Electrofusion is an effective method for fusing two cells\ninto\na hybrid cell, and this method is widely used in immunomedicine, gene\nrecombination, and other related fields. Although cell pairing and\nelectrofusion techniques have been accomplished with microfluidic\ndevices, the purification and isolation of fused cells remains limited\ndue to expensive instruments and complex operations. In this study,\nthrough the optimization of microstructures and electrodes combined\nwith buffer substitution, the entire cell electrofusion process, including\ncell capture, pairing, electrofusion, and precise separation of the\ntargeted fused cells, is achieved on a single chip. The proposed microfluidic\ncell electrofusion achieves an efficiency of 80.2 ± 7.5%, and\ntargeted cell separation could be conveniently performed through the\nstrategic activation of individual microelectrodes via negative dielectrophoresis,\nwhich ensures accurate release of the fused cells with an efficiency\nof up to 91.1 ± 5.1%. Furthermore, the survival rates of the\ncells after electrofusion and release are as high as 94.7 ± 0.6%\nand 91.7 ± 1.2%, respectively. These results demonstrate that\nthe in situ cell electrofusion and separation process did not affect\nthe cell activity. This chip offers integrated multifunctional manipulation\nof cells in situ, and can be applied to multiple fields in the future,\nthus laying the foundation for the field of precise single-cell analysis.