title: 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.
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Deyao
authors.initials: D
authors.lastname: Zhao
authors.affiliation: National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Institute of Microelectronics, Peking University, Beijing 100871, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Huang
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.affiliation: National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Institute of Microelectronics, Peking University, Beijing 100871, China.|Department of Engineering Science and Mechanics, The Pennsylvania State University, State College, PA 16801, USA.
authors.forename: Mengxi
authors.initials: M
authors.lastname: Wu
authors.affiliation: National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Institute of Microelectronics, Peking University, Beijing 100871, China.
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Zhong
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Cheng
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Yidi
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhou
authors.affiliation: National Center for Nanoscience and Technology, Beijing 100190, China.
authors.forename: Zewen
authors.initials: Z
authors.lastname: Wei
authors.affiliation: National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Institute of Microelectronics, Peking University, Beijing 100871, China.
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Li
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Zicai
authors.initials: Z
authors.lastname: Liang
mesh_terms: D000818:Animals; D002460:Cell Line; D018274:Electroporation / Q000295:instrumentation / Q000379:methods; D004912:Erythrocytes / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000295:instrumentation / Q000379:methods
chemical_list: D049452:Green Fluorescent Proteins