Neutrophil pp 43-59 | Cite as

Isolation of Neutrophils from Nonhuman Species

  • Daniel W. Siemsen
  • Liliya N. Kirpotina
  • Natalia Malachowa
  • Igor A. Schepetkin
  • Adeline R. Porter
  • Benfang Lei
  • Frank R. DeLeo
  • Mark T. QuinnEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2087)


The development of new advances in understanding the role of neutrophils in inflammation requires effective procedures for isolating and purifying neutrophils. Methods for isolating human neutrophils are fairly standard, and some are covered in other chapters of this volume and previous editions. However, procedures for isolating neutrophils from nonhuman species used to model human diseases vary from those used in isolating human neutrophils and are not as well developed. Since neutrophils are highly reactive and sensitive to small perturbations, the methods of isolation are important to avoid isolation technique-induced alterations in cell function. We present methods here for reproducibly isolating highly purified neutrophils from large animal models (bovine, equine, ovine), small animal models (murine and rabbit), and nonhuman primates (cynomolgus macaques) and describe optimized details for obtaining the highest cell purity, yield, and viability.

Key words

Inflammation Large animal model Granulocyte Polymorphonuclear leukocyte Cell isolation Flow cytometry Blood Bone marrow 



This work was supported in part by National Institutes of Health IDeA Program COBRE Grant GM110732; the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health; USDA National Institute of Food and Agriculture Hatch project 1009546; and the Montana State University Agricultural Experiment Station.


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Copyright information

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

Authors and Affiliations

  • Daniel W. Siemsen
    • 1
  • Liliya N. Kirpotina
    • 1
  • Natalia Malachowa
    • 2
  • Igor A. Schepetkin
    • 1
  • Adeline R. Porter
    • 2
  • Benfang Lei
    • 1
  • Frank R. DeLeo
    • 2
  • Mark T. Quinn
    • 1
    Email author
  1. 1.Department of Microbiology and ImmunologyMontana State UniversityBozemanUSA
  2. 2.Laboratory of Bacteriology, Rocky Mountain Laboratories, Division of Intramural Research, National Institute of Allergy and Infectious DiseasesNational Institutes of HealthHamiltonUSA

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