Large-Scale mRNA Transfection of Dendritic Cells by Electroporation in Continuous Flow Systems

Dávid Selmeczi, Thomas Steen Hansen, Özcan Met, Inge Marie Svane, Niels Bent Larsen

    Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

    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.
    Original languageEnglish
    Title of host publicationSynthetic mRNA : Production, Introduction Into Cells, and Physiological Consequences
    Number of pages11
    Volume1428
    PublisherSpringer New York
    Publication date2016
    Pages151-61
    Chapter10
    ISBN (Print)978-1-4939-3623-6
    ISBN (Electronic)978-1-4939-3625-0
    DOIs
    Publication statusPublished - 2016
    SeriesMethods in Molecular Biology
    ISSN1064-3745

    Keywords

    • Dendritic cells
    • Electroporation
    • Laminar flow
    • Microfluidics
    • Transfection
    • mRNA

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