Isolation of mouse neutrophils.
Neutrophils are widely used in research to elucidate the mechanisms of inflammation, including such processes as cell adhesion, chemotaxis, superoxide release, production of reactive nitrogen intermediates, and granule exocytosis. This unit presents a protocol for the induction of an enriched exudate of polymorphonuclear leukocytes (PMN) in the peritoneal cavity of mice. Neutrophils are then isolated using continuous gradient centrifugation. This is followed by a protocol for isolating neutrophils from peripheral blood. An alternate method for the purification of PMN from peritoneal exudate cells or peripheral blood by Histopaque density gradient centrifugation is also provided. The cell purification protocol can be modified to obtain nonelicited neutrophils from peritoneal fluid.
Luo
Yi
Y
Harvard Medical School, Boston, Massachusetts.
Dorf
Martin E
ME
Harvard Medical School, Boston, Massachusetts.
D000818:Animals; D001202:Ascitic Fluid / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods; D003973:Diatrizoate; D005362:Ficoll; D051379:Mice; D009504:Neutrophils
D003973:Diatrizoate; D005362:Ficoll; C053816:histopaque