Neutrophil Isolation from Nonhuman Species
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.
KeywordsInflammation Phagocytosis Large animal model Granulocyte Polymorphonuclear leukocyte Cell isolation Flow cytometry Blood Bone marrow
This work was supported in part by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences, National Institutes of Health under grant number GM103500 (D.S., I.A.S., L.K.N., B.L., M.Q.), the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health (N.M., A.R.W., F.R.D.), and the Montana State University Agricultural Experimental Station.