Isolation and Characterization of Murine Bone Marrow-Derived Dendritic Cells

DCs were isolated from mouse bone marrow as previously described 10 (link), 20 (link). Bone marrow mononuclear cells were cultured in RPMI 1640 medium supplemented with 10% FBS, 20 ng/ml recombinant murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF), and 20 ng/ml recombinant murine interleukin (rmIL)-4 (both from Peprotech, Inc) for 7 days. The images of immature bone marrow-derived DCs (BMDCs) were captured using an inverted light microscope (Olympus Corporation). Immature BMDCs were incubated with CD11c MicroBeads UltraPure (Miltenyi Biotec, Inc.) for 20 min at 4°C following the manufacturer's instructions. The positive fraction containing CD11c⁺ cells was analyzed by flow cytometry.

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Fu C., Tian G., Duan J., Liu K., Zhang C., Yan W, & Wang Y. (2021). Therapeutic Antitumor Efficacy of Cancer Stem Cell-Derived DRibble Vaccine on Colorectal Carcinoma. International Journal of Medical Sciences, 18(14), 3249-3260.

Publication 2021

recombinant murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF recombinant murine interleukin (rmIL)-4 inverted light microscope CD11c MicroBeads UltraPure

Bone marrow Bone marrow cells Cells Cultured medium Flow cytometry Granulocyte macrophage colony stimulating factor Light microscope Microbeads Murine Recombinant interleukin 4

Corresponding Organization : Jilin University

Other organizations : VA Palo Alto Health Care System, Stanford University, Minjiang University, Jilin Medical University

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Variable analysis

independent variables

Culture of bone marrow mononuclear cells in RPMI 1640 medium supplemented with 10% FBS, 20 ng/ml recombinant murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF), and 20 ng/ml recombinant murine interleukin (rmIL)-4 for 7 days

dependent variables

Characteristics of immature bone marrow-derived DCs (BMDCs), including their morphology captured using an inverted light microscope
Percentage of CD11c+ cells in the positive fraction analyzed by flow cytometry

control variables

Isolation of DCs from mouse bone marrow as previously described in the referenced studies

controls

No positive or negative controls were explicitly mentioned in the provided information.

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