Neutrophil isolation from nonhuman species

Daniel W. Siemsen, Natalia Malachowa, Igor A. Schepetkin, Adeline R. Whitney, Liliya N. Kirpotina, Benfang Lei, Frank R. Deleo, Mark T. Quinn

Research output: Chapter in Book/Report/Conference proceedingChapter

12 Citations (Scopus)

Abstract

The development of new advances in the understanding of neutrophil biochemistry requires effective procedures for isolating purified neutrophil populations. Although methods for human neutrophil isolation are now standard, similar procedures for isolating neutrophils from many of the nonhuman species used to model human diseases are not as well developed. Since neutrophils are reactive cells, the method of isolation is extremely important to avoid isolation technique-induced alterations in cell function. We present methods here for reproducibly isolating highly purified neutrophils from large animals (bovine, equine, ovine), small animals (murine and rabbit), and nonhuman primates (cynomolgus macaques), and describe optimized details for obtaining the highest cell purity, yield, and viability. We also describe methods to verify phagocytic capacity in the purified cell populations using a flow cytometry-based phagocytosis assay.

Original languageEnglish
Title of host publicationNeutrophil Methods and Protocols
PublisherHumana Press Inc.
Pages19-37
Number of pages19
Volume1124
ISBN (Print)9781627038447
DOIs
Publication statusPublished - 2014
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1124
ISSN (Print)10643745

Fingerprint

Neutrophils
Cell Separation
Macaca
Phagocytosis
Biochemistry
Primates
Population
Horses
Sheep
Flow Cytometry
Rabbits

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Siemsen, D. W., Malachowa, N., Schepetkin, I. A., Whitney, A. R., Kirpotina, L. N., Lei, B., ... Quinn, M. T. (2014). Neutrophil isolation from nonhuman species. In Neutrophil Methods and Protocols (Vol. 1124, pp. 19-37). (Methods in Molecular Biology; Vol. 1124). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-845-4_3

Neutrophil isolation from nonhuman species. / Siemsen, Daniel W.; Malachowa, Natalia; Schepetkin, Igor A.; Whitney, Adeline R.; Kirpotina, Liliya N.; Lei, Benfang; Deleo, Frank R.; Quinn, Mark T.

Neutrophil Methods and Protocols. Vol. 1124 Humana Press Inc., 2014. p. 19-37 (Methods in Molecular Biology; Vol. 1124).

Research output: Chapter in Book/Report/Conference proceedingChapter

Siemsen, DW, Malachowa, N, Schepetkin, IA, Whitney, AR, Kirpotina, LN, Lei, B, Deleo, FR & Quinn, MT 2014, Neutrophil isolation from nonhuman species. in Neutrophil Methods and Protocols. vol. 1124, Methods in Molecular Biology, vol. 1124, Humana Press Inc., pp. 19-37. https://doi.org/10.1007/978-1-62703-845-4_3
Siemsen DW, Malachowa N, Schepetkin IA, Whitney AR, Kirpotina LN, Lei B et al. Neutrophil isolation from nonhuman species. In Neutrophil Methods and Protocols. Vol. 1124. Humana Press Inc. 2014. p. 19-37. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-845-4_3
Siemsen, Daniel W. ; Malachowa, Natalia ; Schepetkin, Igor A. ; Whitney, Adeline R. ; Kirpotina, Liliya N. ; Lei, Benfang ; Deleo, Frank R. ; Quinn, Mark T. / Neutrophil isolation from nonhuman species. Neutrophil Methods and Protocols. Vol. 1124 Humana Press Inc., 2014. pp. 19-37 (Methods in Molecular Biology).
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