THE COMPARATIVE RESPONSES OF HUMAN POLYMORPHONUCLEAR LEUKOCYTES OBTAINED BY COUNTERFLOW CENTRIFUGAL ELUTRIATION AND FICOLL-HYPAQUE DENSITY CENTRIFUGATION .1. RESTING VOLUME, STIMULUS-INDUCED SUPEROXIDE PRODUCTION, AND PRIMARY AND SPECIFIC GRANULE RELEASE

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  • 1 January 1983
    • journal article
    • research article
    • Vol. 102  (5) , 732-742
Abstract
Standard isolation techniques for the human polymorphonuclear leukocyte (PMN) involve sequential exposure of cells to the nonphysiologic environments of detrans, Ficoll-Hypaque (FH) gradient centrifugation and hypotonic conditions. These may be harmful to the recovered PMN. Counterflow centrifugal elutriation (CCE) allows separation of human PNM while the cells are continuously bathed in a physiologic and isotonic buffer. To investigate whether preparative technique may alter PMN activation, PMN obtained by these 2 methods were compared for stimulus-induced superoxide production and release of primary and specific granule contents. Resting PMN volume was also evaluated. PMN obtained using the CCE method were larger and released significantly more superoxide and specific granule contents than PMN obtained by the standard FH technique. The possible origins for these differences are discussed.