DCs were generated as previously described.27 (link) Briefly, bone marrow cells obtained from mouse femurs were cultured in 6-well plates (5×106 cells/well) in culture medium supplemented with 40 ng/mL granulocyte macrophage-colony stimulating factor and 40 ng/mL IL-4 (both from CreaGene, Seongnam, South Korea). After 3 days, the non-adherent cells were removed by gently shaking the dish and then replacing the medium. On day 4, the non-adherent cells were again removed by the same method. On day 6, half the culture medium was replaced with fresh medium. To induce maturation, DCs were exposed to 50 ng/mL interferon (IFN)-γ and 50 ng/mL tumor necrosis factor (TNF)-α (both from PeproTech Inc., Rocky Hill, NJ, USA), or treated with the indicated NPs for 48 h. The DCs were harvested by gentle pipetting on day 8.
Protocol detail
Generating Mature Dendritic Cells
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Bone marrow cells
Cells
Dish
Femurs
Granulocyte macrophage colony stimulating factor
Ifn γ
Interferon
Mouse
Tumor necrosis factor α
Corresponding Organization : Chungbuk National University
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Variable analysis
independent variables
- Exposure to 50 ng/mL interferon (IFN)-γ and 50 ng/mL tumor necrosis factor (TNF)-α
- Treatment with the indicated NPs
- DC maturation
- Bone marrow cells obtained from mouse femurs
- Culture medium supplemented with 40 ng/mL granulocyte macrophage-colony stimulating factor and 40 ng/mL IL-4
- Removal of non-adherent cells on days 3 and 4
- Replacement of half the culture medium with fresh medium on day 6
- Harvesting of DCs on day 8
- Exposure to 50 ng/mL interferon (IFN)-γ and 50 ng/mL tumor necrosis factor (TNF)-α
- Not mentioned
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