Original article| Volume 102, ISSUE 5, P732-742, November 1983

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The comparative responses of human polymorphonuclear leukocytes obtained by counterflow centrifugal elutriation and Ficoll-Hypaque density centrifugation

I. Resting volume, stimulus-induced superoxide production, and primary and specific granule release
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      Standard isolation techniques for the human polymorphonuclear leukocyte (PMN) involve sequential exposure of cells to the nonphysiologic environments of dextran, Ficoll-Hypaque (FH) gradient centrifugation, and hypotonic conditions. It has been suggested that these may be harmful to the recovered PMN. Counterflow centrifugal elutriation (CCE) allows separation of human PMNs while the cells are continuously bathed in a physiologic and isotonic buffer. To investigate whether preparative technique may alter PMN activation, we compared PMNs obtained by these two methods for stimulus-induced superoxide production and release of primary and specific granule contents. Resting PMN volume was also evaluated. We observed that PMNs obtained using the CCE method were larger and released significantly more superoxide and specific granule contents than PMNs obtained by the standard FH technique. The possible origins for these differences are discussed.


      (PMN) (polymorphonuclear leukocyte), (FH) (Ficoll-Hypaque gradient centrifugation technique), (CCE) (counterflow centrifugal elutriation), (FMLP) (N-formyl-methionyl-leucyl-phenylalanine), (O2⨪) (superoxide anion), (EDTA) (ethylenediamine tetracetic acid), (PBS-G) (phosphate-buffered saline with 1 mg/ml glucose), (NEM) (N-ethyl maleimide), (SOD) (superoxide dismutase), (LDH) (lactate dehydrogenase), (DMSO) (dimethyl sulfoxide), (ACD) (acid-citrate-dextrose)
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