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Cell culture plastics with immobilized interleukin-4 for monocyte differentiation

Authors

  • Morten Hansen,

    1. Department of Micro- and Nanotechnology, Technical University of Denmark, DTU-Nanotech, DK-2800 Kongens Lyngby, Denmark
    2. Department of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, Denmark
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  • Gertrud M. Hjortø,

    1. Department of Micro- and Nanotechnology, Technical University of Denmark, DTU-Nanotech, DK-2800 Kongens Lyngby, Denmark
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  • Özcan Met,

    1. Center for Cancer Immune Therapy (CCIT), Departments of Hematology and Oncology, Herlev University Hospital, Denmark
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  • Mogens H. Jakobsen,

    1. Department of Micro- and Nanotechnology, Technical University of Denmark, DTU-Nanotech, DK-2800 Kongens Lyngby, Denmark
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  • Inge M. Svane,

    1. Center for Cancer Immune Therapy (CCIT), Departments of Hematology and Oncology, Herlev University Hospital, Denmark
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  • Niels B. Larsen

    Corresponding author
    1. Department of Micro- and Nanotechnology, Technical University of Denmark, DTU-Nanotech, DK-2800 Kongens Lyngby, Denmark
    • Department of Micro- and Nanotechnology, Technical University of Denmark, DTU-Nanotech, DK-4000 Roskilde, Denmark
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Abstract

Standard cell culture plastic was surface modified by passive adsorption or covalent attachment of interleukin (IL)-4 and investigated for its ability to induce differentiation of human monocytes into mature dendritic cells, a process dose-dependently regulated by IL-4. Covalent attachment of IL-4 proceeded via anthraquinone photochemistry to introduce amine functionalities at the surface followed by coupling of IL-4 through a bifunctional amine-reactive linker. X-ray photoelectron spectroscopy showed that undesirable multilayer formation of the photoactive compound could be avoided by reaction in water instead of phosphate-buffered saline. Passively adsorbed IL-4 was observed to induce differentiation to dendritic cells, but analysis of cell culture supernatants revealed that leakage of IL-4 into solution could account for the differentiation observed. Covalent attachment resulted in bound IL-4 at similar concentrations to the passive adsorption process, as measured by enzyme-linked immunosorbent assays, and the bound IL-4 did not leak into solution to any measurable extent during cell culture. However, covalently bound IL-4 was incapable of inducing monocyte differentiation. This may be caused by IL-4 denaturation or improper epitope presentation induced by the immobilization process, or by biological irresponsiveness of monocytes to IL-4 in immobilized formats. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2011.

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