A high-yield, high-purity elutriation method for preparing human granulocytes demonstrating enhanced experimental lifetimes

Abstract

A counterflow centrifugal elutriation procedure for preparing human granulocytes is described. This method permits isolation of 1-2 X 10(9) granulocytes with a yield of 70% +/- 7% from a unit of whole blood in 3 h. These cells are 99% +/- 1% pure and 95-99% viable. When compared to cells prepared by a conventional procedure employing hypotonic lysis to remove red cells, the elutriated cells show enhanced oxidative responses resulting from enhanced longevity. Other responses including receptor-mediated uptake of chemoattractant were not significantly different from those of conventionally prepared cells after incubation at 37 degrees C prior to stimulation. The population of elutriated granulocytes, however, was significantly more homogeneous and pure than cells prepared conventionally.