Recombinant human gm csf

The UT-7 human cell line was purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). Cell line authentication was performed by DSMZ. The UT-7 cell line was maintained in alpha-MEM (Sigma-Aldrich, Saint Louis, MO, USA) supplemented with 20% fetal bovine serum (Gibco, Gaithersburg, MD, USA), 100 μg/mL streptomycin, 100 units/mL penicillin (Gibco, Gaithersburg, MD, USA), and 2 mM L-glutamine (Gibco, Gaithersburg, MD, USA). Recombinant human GM-CSF (Sigma-Aldrich, Saint Louis, MO, USA) was added to UT-7 cells that require the bioactive protein for survival and proliferation at a final concentration of 5 ng/mL. The cells were cultured in a water-jacketed incubator at 37 °C with 5% CO₂. UT-7 cells were passaged 1:10 twice a week and always 24 h before the experiment. For some of the experiments, the medium was complemented with 80 µM and 100 µM ZnSO₄ (Sigma-Aldrich, Saint Louis, MO, USA).

Cytokines were purchased from Peprotech (Cranbury, NJ, USA): recombinant human IL-4 (#200-04), recombinant human IL-13 (#200-13), recombinant human IL-10 (#200-10), recombinant human M-CSF (#300-25), recombinant human GM-CSF (#300-03), recombinant human IFN-γ (#300-02) and Sigma-Aldrich (St. Louis, MO, USA): LPS (#L3024). Mouse anti-human antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA CD11c FITC (#561355), CD11b FITC (#562793), CD40 BV711 (#563397), CD40 BV510 (#563456), CD80 BV480 (#562432), CD86 BV421 (#562432), MHC-II BV605 (#740408) and from Biolegend (San Diego, CA, USA): CD163 APC (#333610), CD14 PerCp Cy5.5 (#325622), CD206 PE (#321106).