Large-scale isolation of polymorphonuclear leucocytes from bovine blood

A method for the isolation of an enriched population (greater than 96%) of bovine polymorphonuclear leucocytes (PMNs) from 70 liters of blood was developed using a two-step procedure involving separation of the blood, in a packed red blood cell fraction containing the PMNs and a plasma fraction, by continuous flow blood separation. Hypotonic lysis of erythrocytes was then followed by centrifugation at 200 g sedimenting the PMNs. The yield was 93 +/- 30 g, the recovery was 62 +/- 20%, viability was greater than 95%. Since bovine blood can be obtained in unlimited amounts, the procedure described here can be applied to obtain large amounts of bovine PMNs for incubation studies and large-scale purification of intracellular enzymes suitable for biochemical characterization.