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Purification of Human Dendritic Cell Subsets from Peripheral Blood

  • Solana Alculumbre
  • Lucia PattariniEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1423)

Abstract

Blood represents the most accessible source of human dendritic cells (DCs). We present here a method to isolate three DC subtypes, as identified until now, from peripheral blood: plasmacytoid dendritic cells (pDCs), CD141+ myeloid DCs, and CD1c+ myeloid DCs. The method is based on the sequential depletion of non-DCs. First, depletion of granulocytes, erythrocytes, and platelets is obtained by blood centrifugation over a Ficoll gradient. Then, antibodies recognizing non-DCs, combined with magnetic beads, allow enrichment of DCs from peripheral blood mononuclear cells (PBMCs). Finally, enriched DCs are purified and separated into the different subtypes by immunolabeling and fluorescence-activated cell sorting (FACS) using DC-specific surface markers.

DC studies might contribute to the comprehension of human immune processes in physiological and pathological conditions. Human blood DCs targeting might be a useful tool to ameliorate inflammatory diseases and improve vaccination strategies.

Key words

Dendritic cell Human Blood pDC Subtypes 

Notes

Acknowledgments

We are grateful to Vassili Soumelis for his support and suggestions.

We thank Zofia Maciorowski and the Cytometry platform of Curie Institute for help in cell sorting. We thank Cristina Ghirelli and Raphael Zollinger for DC protocol tests and improvement, Carolina Martinez, and Coline Trichot for critical reading and suggestions. S.A. and L.P. were supported by ANR-10-IDEX-0001-02 PSL* and ANR-11-LABX-0043 grants.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.INSERM U932 Immunity and CancerParisFrance
  2. 2.Institut Curie, Centre de rechercheParisFrance

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