A comparative study of neutrophil purification and function.

Several different methods are currently used by numerous laboratories for the isolation of human neutrophils. These methods include partial purification by dextran sedimentation followed by water lysis, and more complete purification procedures utilizing discontinuous density gradients coupled with dextran sedimentation and in some cases hypotonic lysis. Some investigators refrain from using certain purification schemes because certain steps or reagents used in a particular method might adversely affect the functional parameter of the neutrophil they wish to measure. In spite of these concerns there has been no systematic comparison of the functional status of neutrophils prepared by the various methodologies. In this study we have compared 4 commonly used methods of isolation with neutrophil function. The results of this study indicate that while neutrophil yield and purity were determined by isolation procedure, all cells were equivalent with regard to chemotactic performance, ability to degranulate, and ability to produce superoxide and hypochlorous acid.