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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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
Scapini P, Calzetti F, Cassatella MA (1999) On the detection of neutrophil-derived vascular endothelial growth factor (VEGF). J Immunol Methods 232:121–129
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
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
Tecchio C, Micheletti A, Cassatella MA (2014) Neutrophil-derived cytokines: facts beyond expression. Front Immunol 5:508
Nauseef WM (2014) Isolation of human neutrophils from venous blood. Methods Mol Biol 1124:13–18
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
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
Skoog WA, Beck WS (1956) Studies on the fibrinogen, dextran, and phytohemagglutinin methods of isolating leukocytes. Blood 11:436–454
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
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0154-9_3
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0153-2
Online ISBN: 978-1-0716-0154-9
eBook Packages: Springer Protocols