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A High‐Yield, High‐Purity Elutriation Method for Preparing Human Granulocytes Demonstrating Enhanced Experimental Lifetimes
Author(s) -
Tolley James O.,
Omann Geneva M.,
Jesaitis Algirdas J.
Publication year - 1987
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.42.1.43
Subject(s) - elutriation , biology , lysis , yield (engineering) , chromatography , granulocyte , homogeneous , population , incubation , chemotaxis , receptor , immunology , biochemistry , chemistry , materials science , physics , demography , organic chemistry , sociology , metallurgy , thermodynamics
Abstract A counterflow centrifugal elutriation procedure for preparing human granulocytes is described. This method permits isolation of 1‐2 × 10 9 granulocytes with a yield of 70% ± 7% from a unit of whole blood in 3 h. These cells are 99% ±1% pure and 95‐99% viable. When compared to cells prepared by a conventional procedure employing hypotonic lysis to remove red cells, the elutriated cells show enhanced oxidative responses resulting from enhanced longevity. Other responses including receptor‐mediated uptake of chemoattractant were not significantly different from those of conventionally prepared cells after incubation at 37° prior to stimulation. The population of elutriated granulocytes, however, was significantly more homogeneous and pure than cells prepared conventionally.