Advertisement

Dendritic Cell Assays

  • Joanna M. Kreitinger
  • David M. Shepherd
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1803)

Abstract

Generation of dendritic cells from both mouse and human tissues is a valuable technique for downstream immunotoxicological applications. Here, we describe methods for generation of four subsets of dendritic cells from murine bone marrow and three subsets of dendritic cells from human peripheral blood mononuclear cells.

Key words

Dendritic cell Peripheral blood mononuclear cells (PBMCs) Bone marrow-derived dendritic cells (BMDCs) In vitro Cell culture 

Notes

Acknowledgments

This work was supported by NIH grant ES013784 (to D.M.S.) and an AAI Careers in Immunology Fellowship (to J.M.K.).

References

  1. 1.
    Merad M, Sathe P, Helft J, Miller J, Mortha A (2013) The dendritic cell lineage: ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting. Annu Rev Immunol 31:563–604CrossRefPubMedGoogle Scholar
  2. 2.
    Mildner A, Jung S (2014) Development and function of dendritic cell subsets. Immunity 40(5):642–656CrossRefPubMedGoogle Scholar
  3. 3.
    Geissmann F, Auffray C, Palframan R, Wirrig C, Ciocca A, Campisi L, Narni-Mancinelli E, Lauvau G (2008) Blood monocytes: distinct subsets, how they relate to dendritic cells, and their possible roles in the regulation of T-cell responses. Immunol Cell Biol 86(5):398–408CrossRefPubMedGoogle Scholar
  4. 4.
    Simones T, Shepherd DM (2010) Consequences of AhR activation in steady-state dendritic cells. Toxicol Sci 119(2):293–307CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Bankoti J, Rase B, Simones T, Shepherd DM (2010) Functional and phenotypic effects of AhR activation in inflammatory dendritic cells. Toxicol Appl Pharmacol 246(1):18–28CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Gilliet M, Boonstra A, Paturel C, Antonenko S, Xu X-L, Trinchieri G, O'Garra A, Liu Y-J (2002) The development of murine plasmacytoid dendritic cell precursors is differentially regulated by FLT3-ligand and granulocyte/macrophage colony-stimulating factor. J Exp Med 195(7):953–958CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Lutz MB, Kukutsch N, Ogilvie AL, Rößner S, Koch F, Romani N, Schuler G (1999) An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods 223(1):77–92CrossRefPubMedGoogle Scholar
  8. 8.
    Katz SI, Tamaki K, Sachs DH (1979) Epidermal Langerhans cells are derived from cells originating in bone marrow. Nature 282:324–326CrossRefPubMedGoogle Scholar
  9. 9.
    Ohtsuka H, Sakamoto A, Pan J, Inage S, Horigome S, Ichii H, Arima M, Hatano M, Okada S, Tokuhisa T (2011) Bcl6 is required for the development of mouse CD4+ and CD8α+ dendritic cells. J Immunol 186(1):255–263CrossRefPubMedGoogle Scholar
  10. 10.
    Shortman K, Naik SH (2007) Steady-state and inflammatory dendritic-cell development. Nat Rev Immunol 7(1):19–30CrossRefPubMedGoogle Scholar
  11. 11.
    Vremec D, Pooley J, Hochrein H, Wu L, Shortman K (2000) CD4 and CD8 expression by dendritic cell subtypes in mouse thymus and spleen. J Immunol 164(6):2978–2986CrossRefPubMedGoogle Scholar
  12. 12.
    Geissmann F, Prost C, Monnet J-P, Dy M, Brousse N, Hermine O (1998) Transforming growth factor β1, in the presence of granulocyte/macrophage colony-stimulating factor and interleukin 4, induces differentiation of human peripheral blood monocytes into dendritic Langerhans cells. J Exp Med 187(6):961–966CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Dauer M, Obermaier B, Herten J, Haerle C, Pohl K, Rothenfusser S, Schnurr M, Endres S, Eigler A (2003) Mature dendritic cells derived from human monocytes within 48 hours: a novel strategy for dendritic cell differentiation from blood precursors. J Immunol 170(8):4069–4076CrossRefPubMedGoogle Scholar
  14. 14.
    Borras F, Matthews N, Patel R, Navarrete C (2000) Dendritic cells can be successfully generated from CD34+ cord blood cells in the presence of autologous cord blood plasma. Bone Marrow Transplant 26(4):371–376CrossRefPubMedGoogle Scholar
  15. 15.
    Tkachenko N, Wojas K, Tabarkiewicz J, Rolinski J (2005) Generation of dendritic cells from human peripheral blood monocytes-comparison of different culture media. Folia Histochem Cytobiol 43(1):25–30PubMedGoogle Scholar
  16. 16.
    Massa C, Thomas C, Wang E, Marincola F, Seliger B (2015) Different maturation cocktails provide dendritic cells with different chemoattractive properties. J Transl Med 13:175CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Zarif JCHJ, Verdone JE, Campbell SP, Drake CG, Pienta K (2016) A phased strategy to differentiate human CD14+monocytes into classically and alternatively activated macrophages and dendritic cells. BioTechniques 61(1):33–41CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Hulspas R, O’Gorman MRG, Wood BL, Gratama JW, Sutherland DR (2009) Considerations for the control of background fluorescence in clinical flow cytometry. Cytometry 76B:355–364CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Cellular, Molecular, and Microbial Biology Graduate Program, Division of Biological SciencesUniversity of MontanaMissoulaUSA
  2. 2.Department of Biomedical and Pharmaceutical Sciences, Center for Translational MedicineUniversity of MontanaMissoulaUSA

Personalised recommendations