On-chip multiplexed single-cell patterning and controllable intracellular delivery  

作　　者：Zaizai Dong Yanli Jiao Bingteng Xie Yongcun Hao Pan Wang Yuanyuan Liu Junfeng Shi Chandani Chitrakar Stephen Black Yu-Chieh Wang LJames Lee Mo Li Yubo Fan Lingqian Chang 

机构地区：[1]School of Biological Science and Medical Engineering,Beihang University,100083 Beijing,China [2]Institute of Nanotechnology for Single Cell Analysis(INSCA),Beijing Advanced Innovation Center for Biomedical Engineering,Beihang University,100083 Beijing,China [3]College of Agricultural and Life Sciences,University of Florida,Gainesville,FL 32611,USA [4]Center for Reproductive Medicine,Peking University Third Hospital,100191 Beijing,China [5]Ministry of Education Key Laboratory of Micro and Nano Systems for Aerospace,Northwestern Polytechnical University,710072 Xi’an,China [6]Chemical and Biomolecular Engineering Department,Ohio State University,Columbus,OH 43209,USA [7]Department of Biomedical Engineering,University of North Texas,Denton,TX 76207,USA [8]Department of Pharmaceutical Sciences,University of North Texas Health Science Center,Fort Worth,TX 76107,USA

出　　处：《Microsystems & Nanoengineering》2020年第1期1222-1232,共11页微系统与纳米工程（英文）

基　　金：L.C.acknowledges the funding from INSCA at the Beijing Advanced Innovation Center for Biomedical Engineering and Beihang University;start-up funding was received from UNT.

摘　　要：Conventional electroporation approaches show limitations in the delivery of macromolecules in vitro and in vivo.These limitations include low efficiency,noticeable cell damage and nonuniform delivery of cells.Here,we present a simple 3D electroporation platform that enables massively parallel single-cell manipulation and the intracellular delivery of macromolecules and small molecules.A pyramid pit micropore array chip was fabricated based on a silicon wet-etching method.A controllable vacuum system was adopted to trap a single cell on each micropore.Using this chip,safe single-cell electroporation was performed at low voltage.Cargoes of various sizes ranging from oligonucleotides(molecular beacons,22 bp)to plasmid DNA(CRISPR-Cas9 expression vectors,>9 kb)were delivered into targeted cells with a significantly higher transfection efficiency than that of multiple benchmark methods(e.g.,commercial electroporation devices and Lipofectamine).The delivered dose of the chemotherapeutic drug could be controlled by adjusting the applied voltage.By using CRISPR-Cas9 transfection with this system,the p62 gene and CXCR7 gene were knocked out in tumor cells,which effectively inhibited their cellular activity.Overall,this vacuum-assisted micropore array platform provides a simple,efficient,high-throughput intracellular delivery method that may facilitate on-chip cell manipulation,intracellular investigation and cancer therapy.

关 键 词：inhibited VACUUM LIMITATIONS 

分 类 号：TH77[机械工程—仪器科学与技术]