Skip to main content
Book cover

Neutrophil pp 223–233Cite as

Influence of Oxygen on Function and Cholesterol Composition of Murine Bone Marrow-Derived Neutrophils

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

Abstract

During inflammation and infection, invading pathogens as well as infiltrating neutrophils locally consume oxygen and reduce the present oxygen level. Since oxygen is an elementary component of the microenvironment required for cell activity and alters multiple cellular functions, it is important to study neutrophil functionality and phenotype at characteristic pathophysiological oxygen levels that reflect the hypoxic phenotype during infection and inflammation. Here, we describe methods to study murine neutrophils under hypoxic compared to normoxic conditions, including analysis of cholesterol content as a key lipid involved in biological functions.

Key words

  • Hypoxia
  • Normoxia
  • Reactive oxygen species
  • Cholesterol
  • Negative selection
  • Neutrophil isolation

This is a preview of subscription content, access via your institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-0716-0154-9_17
  • Chapter length: 11 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   109.00
Price excludes VAT (USA)
  • ISBN: 978-1-0716-0154-9
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   149.99
Price excludes VAT (USA)
Hardcover Book
USD   219.99
Price excludes VAT (USA)
Fig. 1
Fig. 2
Fig. 3

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Carreau A, El Hafny-Rahbi B, Matejuk A et al (2011) Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. J Cell Mol Med 15:1239–1253

    CAS  CrossRef  Google Scholar 

  2. Lone AG, Atci E, Renslow R et al (2015) Staphylococcus aureus induces hypoxia and cellular damage in porcine dermal explants. Infect Immun 83:2531–2541

    CAS  CrossRef  Google Scholar 

  3. Melican K, Boekel J, Månsson LE et al (2008) Bacterial infection-mediated mucosal signalling induces local renal ischemia as a defense against sepsis. Cell Microbiol 10:1987–1998

    CAS  CrossRef  Google Scholar 

  4. Campbell EL, Bruyninckx WJ, Kelly CJ et al (2014) Transmigrating neutrophils shape the mucosal microenvironment through localized oxygen depletion to influence resolution of inflammation. Immunity 40:66–77

    CAS  CrossRef  Google Scholar 

  5. Schaffer K, Taylor CT (2015) The impact of hypoxia on bacterial infection. FEBS J 282:2260–2266

    CAS  CrossRef  Google Scholar 

  6. Zeitouni NE, Fandrey J, Naim HY et al (2015) Measuring oxygen levels in Caco-2 cultures. Hypoxia 9:53–66

    Google Scholar 

  7. Möllerherm H, Branitzki-Heinemann K, Brogden G et al (2017) Hypoxia modulates the response of mast cells to Staphylococcus aureus infection. Front Immunol 11:541

    CrossRef  Google Scholar 

  8. Branitzki-Heinemann K, Möllerherm H, Völlger L et al (2016) Formation of neutrophil extracellular traps under low oxygen level. Front Immunol 7:518

    CrossRef  Google Scholar 

  9. Brinkmann V, Reichard U, Goosmann C et al (2004) Neutrophil extracellular traps kill bacteria. Science 303:1532–1535

    CAS  CrossRef  Google Scholar 

  10. von Köckritz-Blickwede M, Nizet V (2009) Innate immunity turned inside-out: antimicrobial defense by phagocyte extracellular traps. J Mol Med 87:775–783

    CrossRef  Google Scholar 

  11. Fuchs TA, Abed U, Goosmann C et al (2007) Novel cell death program leads to neutrophil extracellular traps. J Cell Biol 176:231–241

    CAS  CrossRef  Google Scholar 

  12. Stoiber W, Obermayer A, Steinbacher P et al (2015) The role of reactive oxygen species (ROS) in the formation of extracellular traps (ETs) in humans. Biomol Ther 5:702–723

    CAS  Google Scholar 

  13. Neumann A, Brogden G, Jerjomiceva N et al (2014) Lipid alterations in human blood-derived neutrophils lead to formation of neutrophil extracellular traps. Eur J Cell Biol 93:347–354

    CAS  CrossRef  Google Scholar 

  14. Chow OA, von Köckritz-Blickwede M, Bright AT et al (2010) Statins enhance formation of phagocyte extracellular traps. Cell Host Microbe 8:445–454

    CAS  CrossRef  Google Scholar 

  15. Gilk SD, Cockrell DC, Luterbach C et al (2013) Bacterial colonization of host cells in the absence of cholesterol. PLoS Pathog 9:e1003107

    CAS  CrossRef  Google Scholar 

  16. Brogden G, Neumann A, Husein DM et al (2017) Methods to study lipid alterations in neutrophils and the subsequent formation of neutrophil extracellular traps. J Vis Exp (121):54667

    Google Scholar 

  17. Zhao Y, Wu T, Shao S et al (2016) Phenotype, development, and biological function of myeloid-derived suppressor cells. Onco Immunology 5(2)

    CrossRef  Google Scholar 

Download references

Acknowledgments

This work was supported by R2N Project funded by the Federal State of Lower Saxony.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Maren von Köckritz-Blickwede .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Verify currency and authenticity via CrossMark

Cite this protocol

Branitzki-Heinemann, K., Brogden, G., von Köckritz-Blickwede, M. (2020). Influence of Oxygen on Function and Cholesterol Composition of Murine Bone Marrow-Derived Neutrophils. 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_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0154-9_17

  • 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