title: Large-Scale mRNA Transfection of Dendritic Cells by Electroporation in Continuous Flow Systems.
abstract: Electroporation is well established for transient mRNA transfection of many mammalian cells, including immune cells such as dendritic cells used in cancer immunotherapy. Therapeutic application requires methods to efficiently electroporate and transfect millions of immune cells in a fast process with high cell survival. Continuous flow of suspended dendritic cells through a channel incorporating spatially separated microporous meshes with a synchronized electrical pulsing sequence can yield dendritic cell transfection rates of >75 % with survival rates of >90 %. This chapter describes the instrumentation and methods needed for the efficient transfection by electroporation of millions of dendritic cells in one continuous flow process.
authors.affiliation: Department of Micro- and Nanotechnology, DTU Nanotech, Technical University of Denmark, Ørsteds Plads 345B, 2800, Kgs. Lyngby, Denmark.
authors.forename: David
authors.initials: D
authors.lastname: Selmeczi
authors.affiliation: Department of Micro- and Nanotechnology, DTU Nanotech, Technical University of Denmark, Ørsteds Plads 345B, 2800, Kgs. Lyngby, Denmark.
authors.forename: Thomas Steen
authors.initials: TS
authors.lastname: Hansen
authors.affiliation: Center for Cancer Immune Therapy, Department of Hematology, Herlev Hospital, Copenhagen University, Copenhagen, Denmark.|Department of Oncology, Herlev Hospital, Copenhagen University, Copenhagen, Denmark.
authors.forename: Özcan
authors.initials: Ö
authors.lastname: Met
authors.affiliation: Center for Cancer Immune Therapy, Department of Hematology, Herlev Hospital, Copenhagen University, Copenhagen, Denmark.|Department of Oncology, Herlev Hospital, Copenhagen University, Copenhagen, Denmark.
authors.forename: Inge Marie
authors.initials: IM
authors.lastname: Svane
authors.affiliation: Department of Micro- and Nanotechnology, DTU Nanotech, Technical University of Denmark, Ørsteds Plads 345B, 2800, Kgs. Lyngby, Denmark. niels.b.larsen@nanotech.dtu.dk.
authors.forename: Niels B
authors.initials: NB
authors.lastname: Larsen
mesh_terms: D049109:Cell Proliferation; D002470:Cell Survival; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism; D018274:Electroporation / Q000295:instrumentation* / Q000379:methods*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D012333:RNA, Messenger / Q000378:metabolism*; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D049452:Green Fluorescent Proteins
keywords: Dendritic cells; Electroporation; Laminar flow; Microfluidics; Transfection; mRNA