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Abstract
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.
Abbreviations:
(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)To read this article in full you will need to make a payment
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Article info
Publication history
Accepted:
June 27,
1983
Received:
April 5,
1983
Footnotes
☆This work was supported by National Institutes of Health grants PHS R01 AI-10892 and PHS T32 AM 7193, and American Cancer Society grant AMCA IN-46V.
Identification
Copyright
© 1983 Published by Elsevier Inc.