Dendritic cells (DCs) were generated in accordance with our previous work.20 (link) Briefly, bone marrow mononuclear cells were obtained from bone marrow cells after removal of the femurs and tibias of female C3H/HeJ mice. Bone marrow mononuclear cells (1×106/well) were cultured in complete Roswell Park Memorial Institute 1640 medium with 10% fetal bovine serum (Life Technologies, Breda, Netherlands), 10 ng/mL murine granulocyte-macrophage colony-stimulating factor (GM-CSF; eBioscience, San Diego, CA, USA), and 1 ng/mL murine IL-4 (eBioscience) for 5 days. Half the medium with GM-CSF and IL-4 was gently replaced on days 2 and day 4. SCC7, a murine head and neck carcinoma cell line, was cultured in Dulbecco’s Modified Eagle’s Medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/mL penicillin, and 100 mg/mL streptomycin (Invitrogen, Grand Island, NY, USA).
Protocol detail
Dendritic Cell Generation from Mouse Bone Marrow
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fetal bovine serum
murine granulocyte-macrophage colony-stimulating factor (GM-CSF;
murine IL-4
L-glutamine
penicillin
streptomycin
Bone marrow cells
C3h mice
Cell line
Cultured medium
Dendritic cells
Eagle
Female
Femurs
Fetal bovine serum
Glutamine
Gm csf
Head and neck carcinoma
Murine
Penicillin
Streptomycin
Tibias
Corresponding Organization :
Other organizations : Nanjing University, Southeast University, University of Leeds
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Variable analysis
independent variables
- Bone marrow mononuclear cells (1×10^6/well)
- 10 ng/mL murine granulocyte-macrophage colony-stimulating factor (GM-CSF)
- 1 ng/mL murine IL-4
- Dendritic cells (DCs) generated
- Complete Roswell Park Memorial Institute 1640 medium with 10% fetal bovine serum (Life Technologies, Breda, Netherlands)
- Culturing of bone marrow mononuclear cells for 5 days
- Gentle replacement of half the medium with GM-CSF and IL-4 on days 2 and 4
- SCC7, a murine head and neck carcinoma cell line, cultured in Dulbecco's Modified Eagle's Medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/mL penicillin, and 100 mg/mL streptomycin (Invitrogen, Grand Island, NY, USA)
- Positive control: Not specified
- Negative control: Not specified
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