A Flow-Through Cell Electroporation Device for Rapidly and Efficiently Transfecting Massive Amounts of Cells in vitro and ex vivo

Abstract

Continuous cell electroporation is an appealing non-viral approach for genetically transfecting a large number of cells. Yet the traditional macro-scale devices suffer from the unsatisfactory transfection efficiency and/or cell viability due to their high voltage, while the emerging microfluidic electroporation devices is still limited by their low cell processing speed. Here we present a flow-through cell electroporation device integrating large-sized flow tube and small-spaced distributed needle electrode array. Relatively large flow tube enables high flow rate, simple flow characterization and low shear force, while well-organized needle array electrodes produce an even-distributed electric field with low voltage. Thus the difficulties for seeking the fine balance between high flow rate and low electroporation voltage were steered clear. Efficient in vitro electrotransfection of plasmid DNA was demonstrated in several hard-to-transfect cell lines. Furthermore, we also explored ex vivo electroporated mouse erythrocyte as the carrier of RNA. The strong ability of RNA loading and short exposure time of freshly isolated cells jointly ensured a high yield of valid carrier erythrocytes, which further successfully delivered RNA into targeted tissue. Both in vitro and ex vivo electrotransfection could be accomplished at high cell processing speed (20 million cells per minute) which remarkably outperforms previous devices.

The Efficiency of Gene Electrotransfer in Breast-Cancer Cell Lines Cultured on a Novel Collagen-Free 3D Scaffold

“…Considering that the topology and the size of the plasmid DNA vector influence the efficiency of transfection, we focused our analysis on the transfection with a single plasmid construct. In particular, we used the plasmid pEGFP-C3 (4.7 kb), which is a common vector frequently used in model systems of DNA transfection [58]. The voltage pulse parameters used in our study, consisting of applying eight voltage pulses (1300 V/cm, 100 µs long at 1 Hz), were previously demonstrated to assure a high EP efficiency in cells cultured in the same 3D scaffold [50].…”

Methods for protein delivery into cells: from current approaches to future perspectives

“…This allows the application of lower voltage without reducing the electric field strength. This approach minimizes heating and pH changes [55,56]; as a consequence cell viability after electroporation and transfection efficiency are both increased [57][58][59][60][61][62].…”

Evaluation of a Microwave Biosensor for On-Chip Electroporation and Efficient Molecular Delivery Into Mammalian Cells

“…Moreover, the electrically mediated gene transfection is receiving great attention, further motivating the researches working on its miniaturization. Many examples exist in the literature illustrating the on-chip electro-transfection [29], [30], [31]. Finally, the field of Electro Chemo Therapy (ECT), combining cytotoxic agents to an electric field, is rapidly arising in the domain of cancer treatment [32], [33].…”