Technique for Increased Granulocyte Recovery from Human Whole Blood by Counterflow Centrifugation Elutriation

Abstract

Human granulocytes were isolated from 120 ml of whole blood by a modified counterflow centrifugation-elutriation (CCE) technique. Overall recovery of isolated granulocytes averaged 2.82 +/- 0.25 x 10(8) cells or 77 per cent yield from whole blood with 96 per cent granulocyte purity and 4 per cent mononuclear leukocytes. The granulocyte fraction was assayed in vitro to determine chemotactic response, stimulated oxygen consumption in the presence of latex beads, bactericidal capacity, and enzyme activities. Cellular integrity was determined by scanning and transmission electron microscopy as well as by cell volume analysis. The data suggest that granulocytes isolated by CCE suffered no discernible loss of function or morphologic damage. The granulocytes are free of platelets and most mononuclear leukocytes and erythrocytes, and have not been exposed to sedimenting agents or surface adhesive agents such as dextran, nylon or glass wool.