A method for isolating neutrophils from moderate volumes of human blood.

The Flow Laboratories procedure for isolating polymorphonuclear leukocytes (PMNs) from human blood whereby whole blood is centrifuged through Mono-Poly resolving medium (MPRM) has been modified. Using this modification, as much as 30 ml of whole blood could be processed in a single centrifuge tube, and erythrocyte contamination of the final PMN suspension was reduced significantly. PMN suspensions were composed of 96% neutrophils, and neutrophil recovery was 42-55% depending upon the volume of blood processed. Cell viability was 98% as determined by trypan blue exclusion. Greater than 99% of the neutrophils were spherical suggesting that the neutrophils were not primed during isolation.