Cell culture plastics with immobilized interleukin-4 for monocyte differentiation.

Morten Hansen, Gertrud Malene Hjortø, Ozcan Met, Mogens Havsteen Jakobsen, Inge M Svane, Niels Bent Larsen

    Research output: Contribution to journalJournal articleResearchpeer-review

    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.
    Original languageEnglish
    JournalJournal of Biomedical Materials Research. Part A
    Volume96
    Issue number2
    Pages (from-to)372-383
    ISSN1549-3296
    DOIs
    Publication statusPublished - 2011

    Keywords

    • Immobilization process
    • Bifunctional
    • Cells
    • surface modification
    • monocyte
    • Cell culture
    • Amine functionality
    • Enzyme linked immunosorbent assay
    • X ray photoelectron spectroscopy
    • IL-4
    • Dendritic cells
    • Animal cell culture
    • cell culture plastic
    • Saline water
    • Standard cell
    • Covalent attachment
    • Phosphate-buffered salines
    • Supernatants
    • Body fluids
    • differentiation
    • Adsorption process
    • Multilayer formation
    • Human monocytes
    • Photoactive compounds
    • Ketones
    • Electric batteries
    • Interleukin-4
    • Adsorption
    • Surface-modified

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