Abstract
Electroporation is the most widely used transfection method for delivery of cell-impermeable molecules into cells. We developed a novel gene transfection method, water-in-oil (W/O) droplet electroporation, using dielectric oil and an aqueous droplet containing mammalian cells and transgene DNA. When a liquid droplet suspended between a pair of electrodes in dielectric oil is exposed to a DC electric field, the droplet moves between the pair of electrodes periodically and droplet deformation occurs under the intense DC electric field. During electrostatic manipulation of the droplet, the local intense electric field and instantaneous short circuit via the droplet due to droplet deformation facilitate gene transfection. This method has several advantages over conventional transfection techniques, including co-transfection of multiple transgene DNAs into even as few as 103 cells, transfection into differentiated neural cells, and the capable establishment of stable cell lines. In addition, there have been improvements in W/O droplet electroporation electrodes for disposable 96-well plates making them suitable for concurrent performance without thermal loading by a DC electric field. This technique will lead to the development of cell transfection methods for novel regenerative medicine and gene therapy.
Highlights
A variety of cell-impermeable molecules, such as DNA, RNA, proteins, antibodies, and dyes, have been delivered into cells in a wide range of fields, such as life science, medicine, pharmacy, and agriculture
We demonstrated a quite different electroporation method based on electrostatic manipulation of W/O droplets in a direct current (DC) electric field
We developed a novel gene transfection method, water-in-oil (W/O) droplet electroporation, using dielectric oil and an aqueous droplet containing mammalian cells and transgene DNA
Summary
A variety of cell-impermeable molecules, such as DNA, RNA, proteins, antibodies, and dyes, have been delivered into cells in a wide range of fields, such as life science, medicine, pharmacy, and agriculture. Transfection, which is a fundamental technique used to deliver nucleic acids into mammalian cells, is widely used for experimental and therapeutic purposes. Viral vector, electroporation, lipofection, particle gun, and sonoporation methods have been used for transfection. These methods have both advantages and disadvantages with regard to delivery efficiency, viability, running costs, etc. The development of safer, more effective, and low-cost novel transfection methods is required. Electroporation generates transient membrane pores when the transmembrane potential exceeds a critical threshold value, altering
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