Transfection of cells using flow-through electroporation based on constant voltage
Abstract
Electroporation is a high-efficiency and low-toxicity physical gene transfer method. Classical electroporation protocols are limited by the small volume of cell samples processed (less than 107 cells per reaction) and low DNA uptake due to partial permeabilization of the cell membrane. Here we describe a flow-through electroporation protocol for continuous transfection of cells, using disposable devices, a syringe pump and a low-cost power supply that provides a constant voltage. We show transfection of cell samples with rates ranging from 40 mu l min(-1) to 20 ml min(-1) with high efficiency. By inducing complex migrations of cells during the flow, we also show permeabilization of the entire cell membrane and markedly increased DNA uptake. The fabrication of the devices takes 1 d and the flow-through electroporation typically takes 1-2 h.
Keywords
VIRUS VECTOR INTEGRATION; GENE-TRANSFER; STEM-CELLS; MICROFLUIDIC ELECTROPORATION; MAMMALIAN-CELLS; CYSTIC-FIBROSIS; DNA DELIVERY; IN-VIVO; THERAPY; SCID-X1
DOI
10.1038/nprot.2011.360
Citation
Nature Protocols 6, 1192–1208 (2011)
Date of this Version
8-2011
Recommended Citation
Geng, Tao; Zhan, Yihong; Wang, Jun; and Lu, Chang, "Transfection of cells using flow-through electroporation based on constant voltage" (2011). Birck and NCN Publications. Paper 979.
http://dx.doi.org/10.1038/nprot.2011.360