Journal article
Cell culture plastics with immobilized interleukin-4 for monocyte differentiation
Department of Micro- and Nanotechnology, Technical University of Denmark1
Department of Micro- and Nanotechnology, Technical University of Denmark2
Institut for Molekylærbiologi og Genetik3
Section of Surgery and Internal Medicine, Department of Clinical Medicine, Faculty of Health and Medical Sciences, Københavns Universitet4
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.
Language: | English |
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Year: | 2011 |
Pages: | 372-83 |
ISSN: | 15524965 and 15493296 |
Types: | Journal article |
DOI: | 10.1002/jbm.a.32986 |
ORCIDs: | Svane, Inge M |