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Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation
Published on: July 8, 2011
Large-Scale mRNA Transfection of Dendritic Cells by Electroporation in Continuous Flow Systems
David Selmeczi1, Thomas Steen Hansen1, Özcan Met2,3
1Department of Micro- and Nanotechnology, DTU Nanotech, Technical University of Denmark, Ørsteds Plads 345B, 2800, Kgs. Lyngby, Denmark.
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.

