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Isolation of Human Neutrophils from Venous Blood

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Neutrophil

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2087))

Abstract

Venous blood provides a ready source of large numbers of unstimulated granulocytes and mononuclear cells. Exploiting the differences in the relative densities of the leukocytes circulating in venous blood, one can separate leukocytes from erythrocytes as well as isolate the individual leukocyte populations in high purity for use in ex vivo studies. For selected functional studies, such as transcriptional analysis or cytokine quantitation, addition of an immunomagnetic negative selection step to the standard isolation protocol can yield highly purified human neutrophils.

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References

  1. Böyum A (1968) Isolation of mononuclear cells and granulocytes from human blood: isolation of mononuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl 97:77–89

    PubMed  Google Scholar 

  2. Scapini P, Calzetti F, Cassatella MA (1999) On the detection of neutrophil-derived vascular endothelial growth factor (VEGF). J Immunol Methods 232:121–129

    Article  CAS  Google Scholar 

  3. Tamassia N, Cassatella MA, Bazzoni F (2014) Fast and accurate quantitative analysis of cytokine gene expression in human neutrophils. Methods Mol Biol 1124:451–467

    Article  CAS  Google Scholar 

  4. Tamassia N, Zimmermann M, Castellucci M et al (2013) Cutting edge: an inactive chromatin configuration at the IL-10 locus in human neutrophils. J Immunol 190:1921–1925

    Article  CAS  Google Scholar 

  5. Tecchio C, Micheletti A, Cassatella MA (2014) Neutrophil-derived cytokines: facts beyond expression. Front Immunol 5:508

    Article  Google Scholar 

  6. Nauseef WM (2014) Isolation of human neutrophils from venous blood. Methods Mol Biol 1124:13–18

    Article  CAS  Google Scholar 

  7. Zimmermann M, Aguilera FB, Castellucci M et al (2015) Chromatin remodelling and autocrine TNFα are required for optimal interleukin-6 expression in activated human neutrophils. Nat Commun 6:6061

    Article  CAS  Google Scholar 

  8. Zimmermann M, Arruda-Silva F, Bianchetto-Aguilera F et al (2016) IFNalpha enhances the production of IL-6 by human neutrophils activated via TLR8. Sci Rep 6:19674

    Article  CAS  Google Scholar 

  9. Skoog WA, Beck WS (1956) Studies on the fibrinogen, dextran, and phytohemagglutinin methods of isolating leukocytes. Blood 11:436–454

    Article  CAS  Google Scholar 

  10. Stie J, Jesaitis AJ (2007) Reorganization of the human neutrophil plasma membrane is associated with functional priming: implications for neutrophil preparations. J Leuk Biol 81:672–685

    Article  CAS  Google Scholar 

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Correspondence to William M. Nauseef .

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Kremserova, S., Nauseef, W.M. (2020). Isolation of Human Neutrophils from Venous Blood. In: Quinn, M., DeLeo, F. (eds) Neutrophil. Methods in Molecular Biology, vol 2087. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0154-9_3

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  • DOI: https://doi.org/10.1007/978-1-0716-0154-9_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0153-2

  • Online ISBN: 978-1-0716-0154-9

  • eBook Packages: Springer Protocols

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