Isolation and Differentiation of Murine Bone Marrow-Derived Dendritic Cells

Bone marrow-derived progenitor cells were isolated from femurs, tibiae and hip bones of non-transgenic C57BL/6J mice according to established protocols (30 (link)). Progenitor cells were cultivated in RPMI-1640 with 10% (v/v) FBS (heat-inactivated), GlutaMAX supplement (1:100 from stock, Gibco, Thermo Scientific), 50 μM β-mercaptoethanol (Sigma) and 200 U/ml murine Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF, PeproTech) at 0.2 Mio. cells/ml. After 9 days of incubation at 37°C and 5% CO₂, progenitor cells completely differentiated into immature bone marrow-derived dendritic cells (BM-DCs).

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Gericke C., Mallone A., Engelhardt B., Nitsch R.M, & Ferretti M.T. (2020). Oligomeric Forms of Human Amyloid-Beta(1–42) Inhibit Antigen Presentation. Frontiers in Immunology, 11, 1029.

Publication 2020

GlutaMAX supplement β-mercaptoethanol murine Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF,

Cells Cells bone marrow Dendritic Femurs Gm csf Hip bones Mercaptoethanol Murine Progenitor cells Supplement Tibiae Transgenic mice

Corresponding Organization : University of Zurich

Other organizations : University of Bern

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Variable analysis

independent variables

Isolation of bone marrow-derived progenitor cells from femurs, tibiae and hip bones of non-transgenic C57BL/6J mice

dependent variables

Differentiation of progenitor cells into immature bone marrow-derived dendritic cells (BM-DCs)

control variables

Cultivation of progenitor cells in RPMI-1640 medium with 10% (v/v) FBS (heat-inactivated), GlutaMAX supplement (1:100 from stock), 50 μM β-mercaptoethanol, and 200 U/ml murine Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF)
Incubation of progenitor cells at 37°C and 5% CO2 for 9 days

controls

The authors do not explicitly mention any positive or negative controls in the provided information.

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