Rapid method for the isolation of neutrophils in high yield without the use of dextran or density gradient polymers.
A simple, rapid and economical method for the isolation of polymorphonuclear leucocytes (PMNs) from whole blood is compared with dextran and dextran/Lymphoprep gradient techniques. The method eliminates the use of dextran and density gradient polymers such as Ficoll which have been shown to affect PMNs adversely. The technique is based on the lysis of red cells with isotonic ammonium chloride solution followed by differential centrifugation to separate the PMNs. This method gave a PMN yield of 73% (SD +/- 3.5) and a purity of 78% (SD +/- 2.5). Both morphology and functional activity were preserved, as assessed by bacterial phagocytosis and killing, chemotaxis, polarising response, superoxide production and adherence. In contrast, the dextran and dextran/Lymphoprep techniques gave yields of 50% and 15% with purities of 78% and 91% respectively. In a series of 14 PMN isolations, the differential centrifugation method gave an average yield of 63% with an average purity of 83%.
Eggleton
P
P
Department of Biochemistry, Royal Free Hospital School of Medicine, University of London, U.K.
Gargan
R
R
Fisher
D
D
D000643:Ammonium Chloride / Q000494:pharmacology; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D003911:Dextrans / Q000494:pharmacology; D006801:Humans; D009504:Neutrophils / Q000502:physiology*
D003911:Dextrans; D000643:Ammonium Chloride