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Isolation of Mouse Dendritic Cell Subsets and Macrophages from the Skin

  • Camille Malosse
  • Sandrine HenriEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1423)

Abstract

The improvement of dendritic cell subset isolation from tissues and the use of appropriate surface markers allowed to decipher their heterogeneity but also allowed to unravel some specific functions that are valuable for vaccine design as well as for a better understanding of the in situ pathophysiology upon infection. Here, we describe the procedures to extract those cells from the skin and to analyze them by flow cytometry using a combination of appropriate surface markers allowing further transcriptomic analysis and functional assays.

Key words

Skin Epidermis Dermis Dispase Collagenase 4 Percoll Fc Block Monoclonal antibodies Flow cytometry 

Notes

Acknowledgment

This work is supported by ANR JCJC Skin DCs.

References

  1. 1.
    Steinman RM, Cohn ZA (1973) Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution. J Exp Med 137(5):1142–1162CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Brasel K, De Smedt T, Smith JL, Maliszewski CR (2000) Generation of murine dendritic cells from flt3-ligand-supplemented bone marrow cultures. Blood 96(9):3029–3039PubMedGoogle Scholar
  3. 3.
    Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S, Muramatsu S, Steinman RM (1992) Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med 176(6):1693–1702CrossRefPubMedGoogle Scholar
  4. 4.
    Robbins SH, Walzer T, Dembele D, Thibault C, Defays A, Bessou G, Xu H, Vivier E, Sellars M, Pierre P, Sharp FR, Chan S, Kastner P, Dalod M (2008) Novel insights into the relationships between dendritic cell subsets in human and mouse revealed by genome-wide expression profiling. Genome Biol 9(1):R17CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Xu Y, Zhan Y, Lew AM, Naik SH, Kershaw MH (2007) Differential development of murine dendritic cells by GM-CSF versus Flt3 ligand has implications for inflammation and trafficking. J Immunol 179(11):7577–7584CrossRefPubMedGoogle Scholar
  6. 6.
    Malissen B, Tamoutounour S, Henri S (2014) The origins and functions of dendritic cells and macrophages in the skin. Nat Rev Immunol 14(6):417–428CrossRefPubMedGoogle Scholar
  7. 7.
    Poulin LF, Henri S, de Bovis B, Devilard E, Kissenpfennig A, Malissen B (2007) The dermis contains langerin + dendritic cells that develop and function independently of epidermal Langerhans cells. J Exp Med 204(13):3119–3131CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Tamoutounour S, Guilliams M, Montanana Sanchis F, Liu H, Terhorst D, Malosse C, Pollet E, Ardouin L, Luche H, Sanchez C, Dalod M, Malissen B, Henri S (2013) Origins and functional specialization of macrophages and of conventional and monocyte-derived dendritic cells in mouse skin. Immunity 39(5):925–938CrossRefPubMedGoogle Scholar
  9. 9.
    Crozat K, Tamoutounour S, Vu Manh TP, Fossum E, Luche H, Ardouin L, Guilliams M, Azukizawa H, Bogen B, Malissen B, Henri S, Dalod M (2011) Cutting edge: expression of XCR1 defines mouse lymphoid-tissue resident and migratory dendritic cells of the CD8alpha+ type. J Immunol 187(9):4411–4415CrossRefPubMedGoogle Scholar
  10. 10.
    Henri S, Poulin LF, Tamoutounour S, Ardouin L, Guilliams M, de Bovis B, Devilard E, Viret C, Azukizawa H, Kissenpfennig A, Malissen B (2010) CD207+ CD103+ dermal dendritic cells cross-present keratinocyte-derived antigens irrespective of the presence of Langerhans cells. J Exp Med 207(1):189–206CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Guilliams M, Crozat K, Henri S, Tamoutounour S, Grenot P, Devilard E, de Bovis B, Alexopoulou L, Dalod M, Malissen B (2010) Skin-draining lymph nodes contain dermis-derived CD103(-) dendritic cells that constitutively produce retinoic acid and induce Foxp3(+) regulatory T cells. Blood 115(10):1958–1968CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Centre d’Immunologie de Marseille-Luminy (CIML)UM2 Aix-Marseille UniversitéMarseille Cedex 9France
  2. 2.INSERM U1104Marseille Cedex 9France
  3. 3.CNRS UMR7280Marseille Cedex 9France

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