Sterile isolation of polymorphonuclear leukocytes from large blood volumes

A method for preparation of pure polymorphonuclear leukocyte populations from large blood volumes is described. Methyl cellulose sedimentation and hypotonic haemolysis resulted in an efficient removal of erythrocytes. Subsequent separation with sodium metrizoate/Ficoll (density 1.077) gave rise to a preparation containing 95% polymorphonuclear leukocytes with a recovery of 56% and complete exclusion of red cell ghosts. The granulocyte function expressed by chemotaxis to the standard attractant casein and by trypan blue exclusion was not influenced by the separation procedure, which may be applied prior to granulocyte labeling for studies of cell kinetics and inflammatory disease.