Abstract
Advances in the understanding of neutrophil biochemistry require the development of 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. Because 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 (cow, horse, sheep) and small (mouse, rabbit) animal models 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 language | English |
|---|---|
| Pages (from-to) | 21-34 |
| Number of pages | 14 |
| Journal | Methods in Molecular Biology |
| Volume | 412 |
| Publication status | Published - 2007 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Genetics
- Molecular Biology
Cite this
Neutrophil isolation from nonhuman species. / Siemsen, Daniel W.; Schepetkin, Igor A.; Kirpotina, Liliya N.; Lei, Benfang; Quinn, Mark T.
In: Methods in Molecular Biology, Vol. 412, 2007, p. 21-34.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Neutrophil isolation from nonhuman species.
AU - Siemsen, Daniel W.
AU - Schepetkin, Igor A.
AU - Kirpotina, Liliya N.
AU - Lei, Benfang
AU - Quinn, Mark T.
PY - 2007
Y1 - 2007
N2 - Advances in the understanding of neutrophil biochemistry require the development of 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. Because 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 (cow, horse, sheep) and small (mouse, rabbit) animal models 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.
AB - Advances in the understanding of neutrophil biochemistry require the development of 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. Because 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 (cow, horse, sheep) and small (mouse, rabbit) animal models 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=45549104352&partnerID=8YFLogxK
M3 - Article
VL - 412
SP - 21
EP - 34
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -