Granulocyte Separation by Modified Centrifugal Elutriation System

Abstract

The Beckman centrifugal elutriation system was modified for a 1-step separtion of granulocytes from whole human blood. A special valve for reversing the direction of flow permits the collection of concentrated granulocytes in small volumes and allows unclogging of the separation chamber should cell packing occur during centrifugal elutriation. Blood diluted 10-fold prevents coagulation, reduces the cell trauma and makes separation more efficient. Maintenance of the precise flow rates by visual monitoring of the granulocyte level in the chamber during elutriation, when the chamber begins to clear of red blood cells, is essential for optimal separation. Granulocytes isolated at rotor speed of 2500 rpm are indistinguishable in their membrane integrity, bactericidal and phagocytic abilities from those collected by dextran-sedimentation. Cell yields close to 100%, high purity, minimal cell traumatization and short processing time make this technique highly effective in collecting granulocytes for clinical and laboratory studies.