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Isolation of Mouse Thymic Dendritic Cells

  • Protocol
Dendritic Cell Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 64))

Abstract

The method described in this chapter for the isolation of mouse thymic dendritic cells (DC) is an optimization of our previously published methods (1,2) and involves the following major steps:

  1. 1.

    Enzymatic digestion of thymic fragments with collagenase and DNase.

  2. 2.

    Separation of a very-low-density cell fraction (VLDF).

  3. 3.

    Magnetic depletion of T-lineage cells, B cells, macrophages, and granulocytes.

  4. 4.

    Positive selection of DCs by magnetic cell sorting (MACS).

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References

  1. Vremec, D., Zorbas, M., Scollay, R. Saunders, D. J., Ardavin, C. F., and Shortman, K. (1992). The surface phenotype of dendritic cells purified from mouse thymus and spleen: Investigation of the CD8 expression by a subpopulationv of dendritic cells. J. Exp.Med. 176, 47–58.

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  3. Ardavin C., Wu, L., Li, C.-L., and Shortman, K. (1993). Thymic dendritic cells and T cells develop simultaneously in the thymus from a common precursor population. Nature 362, 761–763.

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  4. Ardavin, C. (1997). Thymic dendritic cells. Immunol Today 18, 350–361.

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  5. Wu, L., Vremec, D., Ardavin, C., Winkel, K., Suss, G., Georgiou, H., Maraskovsky E., Cook, W., and Shortman, K. (1995). Mouse thymus dendritic cells: kinetics of development and changes in surface markers during maturation. Eur. J. Immunol. 25, 418–425.

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Author information

Authors and Affiliations

  1. Department of Cell Biology, Complutense University, Madrid, Spain

    Fabienne Anjuère & Carlos ArdavÍn

Authors
  1. Fabienne Anjuère
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  2. Carlos ArdavÍn
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Editor information

Editors and Affiliations

  1. Department of Hematology, University College Hospital London, UK

    Stephen P. Robinson MD, PhD

  2. Antigen Presentation Research Group, Imperial College School of Medicine, at Northwick Park Hospital, Harrow, Middlesex, UK

    Andrew J. Stagg PhD

  3. Professor of Immunopathology, Imperial College School of Medicine, London

    Stella C. Knight PhD

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© 2001 Humana Press Inc.

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Anjuère, F., ArdavÍn, C. (2001). Isolation of Mouse Thymic Dendritic Cells. In: Robinson, S.P., Stagg, A.J., Knight, S.C. (eds) Dendritic Cell Protocols. Methods in Molecular Medicine™, vol 64. Humana Press. https://doi.org/10.1385/1-59259-150-7:23

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  • DOI: https://doi.org/10.1385/1-59259-150-7:23

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-584-3

  • Online ISBN: 978-1-59259-150-3

  • eBook Packages: Springer Protocols

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