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mRNA-electroporated mature dendritic cells retain transgene expression, phenotypical properties and stimulatory capacity after cryopreservation

Leukemia volume 16, pages 1324–1330 (2002)Cite this article

Abstract

Genetically modified dendritic cells (DC) are increasingly used in vitro to activate cytotoxic T lymphocyte (CTL) immune responses. Because T cell activation protocols consist of multiple restimulation cycles of peripheral blood lymphocytes with antigen-loaded mature DC, continuous generation of DC is needed throughout the experiment. Therefore, cryopreservation of DC loaded with antigen is a valuable alternative for weekly generation and modification of DC. Recently, we described an antigen loading method for DC based on electroporation of defined tumor antigen mRNA. In this study, we demonstrate that mRNA-electroporated DC can efficiently be prepared for cryopreservation. Using an optimized maturation and freezing protocol after mRNA electroporation, we obtained high transgene-expressing viable mature DC. In addition, we showed that these modified cryopreserved DC retain stimulatory capacity in an influenza model system. Therefore, cryopreservation of mature mRNA-electroporated DC is a useful method for continuous availability of antigen-loaded DC throughout T cell activation experiments.

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Acknowledgements

This work was supported by grant Nos 3.0109.96, G.0157.99 and G.0313.01 of the Fund for Scientific Research – Flanders, Belgium (FWO), by grants of the Scientific Committee of the Fortis Bank (FB)-financed Cancer Research and of the Belgian Federation against Cancer (BFK). PP, NC, AVD are holders of a PhD fellowship from the Flemish Institute for Science and Technology (IWT). VFIVT is a postdoctoral fellow of the FWO.

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Authors and Affiliations

  1. Laboratory of Experimental Hematology, Antwerp University Hospital (UZA), University of Antwerp (UIA), Edegem, Belgium

    P Ponsaerts, VFI Van Tendeloo, N Cools, A Van Driessche, G Nijs, M Lenjou, DR Van Bockstaele & ZN Berneman

  2. Laboratory of Cancer Research and Clinical Oncology, University of Antwerp (UIA), Antwerpen, Belgium

    F Lardon

  3. Antwerp Blood Transfusion Center, Edegem, Belgium

    G Mertens

  4. Laboratory of Molecular Genetics, University of Antwerp (UIA), Antwerpen, Belgium

    C Van Broeckhoven

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  1. P Ponsaerts
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Ponsaerts, P., Van Tendeloo, V., Cools, N. et al. mRNA-electroporated mature dendritic cells retain transgene expression, phenotypical properties and stimulatory capacity after cryopreservation. Leukemia 16, 1324–1330 (2002). https://doi.org/10.1038/sj.leu.2402511

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