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

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1428; p. 151
• Main Authors: Selmeczi, David, Hansen, Thomas Steen, Met, Özcan, Svane, Inge Marie, Larsen, Niels B
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Cell Proliferation; Cell Survival; Dendritic Cells - cytology; Dendritic Cells - metabolism; Electroporation - instrumentation; Electroporation - methods; Gene Transfer Techniques; Green Fluorescent Proteins - genetics; Humans; RNA, Messenger - metabolism; Transfection; Electroporation; mRNA; Transfection; Dendritic cells; Laminar flow; Microfluidics
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-3625-0_10

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.