A high-yield, high-purity elutriation method for preparing human granulocytes demonstrating enhanced experimental lifetimes.
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.
D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002498:Centrifugation; D006098:Granulocytes / Q000166:cytology*; D006801:Humans; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000097:blood; D009504:Neutrophils / Q000166:cytology; D013481:Superoxides / Q000097:blood; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D013755:Tetradecanoylphorbol Acetate