Dendritic Cell Generation from Human and Murine Sources

Human monocyte-derived dendritic cells (mDC) were generated from CD14⁺ monocytes isolated from human PBMC by magnetic bead separation (Miltenyi) and were differentiated by culturing for 6-days in RPMI1640 (Lonza) 5% FCS (Biosera) supplemented with 20 ng/ml GM-CSF (Life Technologies) and 4 ng/ml IL-4 (BD Biosciences) as described previously (8 (link)). CGD donors were recruited at the Royal Free Hospital, London and age and sex matched to healthy donors recruited at the University of Cambridge, in accordance with ethical rules set out by each institution. Human CD1c⁺ DC were isolated from PBMC by magnetic bead separation (Miltenyi). Murine bone marrow-derived dendritic cells (BMDC) were generated by culturing cells isolated from bone marrow of wildtype (TSLPR^+/+) and TSLPR knockout (TSLPR^−/−) BALB/c mice for 7-days in RPMI1640 10% FCS supplemented with 5% X63 conditioned media and 10 ng/ml IL-4 (Peprotech).

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Elder M.J., Webster S.J., Fitzmaurice T.J., Shaunak A.S., Steinmetz M., Chee R., Mallat Z., Cohen E.S., Williams D.L., Gaston J.S, & Goodall J.C. (2019). Dendritic Cell-Derived TSLP Negatively Regulates HIF-1α and IL-1β During Dectin-1 Signaling. Frontiers in Immunology, 10, 921.

Publication 2019

magnetic bead separation RPMI1640 FCS GM-CSF IL-4

Balb c mice Bone marrow Bone marrow cells Cd1c Conditioned media Dendritic Dendritic cells Donors Gm csf Human Monocytes Murine Sex healthy

Corresponding Organization : University of Cambridge

Other organizations : AstraZeneca (United Kingdom), Paris Cardiovascular Research Center, Inserm, The Royal Free Hospital, University College London, East Tennessee State University

Top 5 similar protocols

Variable analysis

independent variables

TSLPR knockout (TSLPR^-/-) BALB/c mice
Wildtype (TSLPR^+/+) BALB/c mice

dependent variables

Murine bone marrow-derived dendritic cells (BMDC)

control variables

Culturing cells isolated from bone marrow for 7-days in RPMI1640 10% FCS supplemented with 5% X63 conditioned media and 10 ng/ml IL-4 (Peprotech)

positive controls

Wildtype (TSLPR^+/+) BALB/c mice

negative controls

TSLPR knockout (TSLPR^-/-) BALB/c mice

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