Flow Micropillar Array Electroporation to Enhance Size Specific Transfection to A Large Population of Cells

Abstract Despite serving as a popular non-viral delivery approach, electroporation carries several drawbacks in its current configurations. We developed a F low M icropillar-array E lectroporation (FME) system to wisely regulate an important transmembrane-determining factor, namely cell size variations among individual cells, to achieve effective transfection. In FME, cells flow through a slit-type microfluidic channel on which carbon electrodes with well-patterned micropillar array texture are integrated as the top and bottom wall. Gravity helps bring cells to the micropillar array surface so that the permeable area on cells in different size populations is specified by their size regardless their random location fact. Without sacrificing cell viability, we demonstrate this FME concept by delivering DNA plasmids to several mammalian cell lines with obvious transfection enhancement when compared to a commercial system (K562: 3.0 folds; A549: 3.3 folds; HeLa: 1.8 folds, COS7: 1.7 folds; 293T: 2.9 folds; mES: 2.5 folds). Moreover, carbon-based electrodes are less expensive, more durable, and convenient for integration with a microfluidic setup which enables rapid and massive transfection capability that many therapeutic application needs. The success of FME may benefit many emerging biological studies and clinical practice that requires effective transfection to a large population of cells in limited processing time.