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Purification and functional evaluation of mature neutrophils from human
bone marrow
RL Berkow and RW Dodson
Human myeloid maturation proceeds within the bone marrow and results in a
mature neutrophil that is released into the peripheral circulation.
Previous reports have indicated that neutrophils from bone marrow
demonstrate decreased adherence, impaired phagocytosis, and decreased
nitroblue tetrazolium dye reduction when stimulated. Due to lack of a
suitable method for isolating purified bone marrow neutrophils, these
studies have been performed by microscopic techniques. We now report a
method for isolating 1 X 10(8) neutrophils [bands plus polymorphonuclear
leukocytes (PMNs)] from 10 mL of bone marrow aspirate sample. By means of a
discontinuous Percoll-gradient centrifugation through densities of 1.085,
1.095, and 1.10 g/mL a leukocyte-rich suspension of bone marrow can be
separated into three leukocyte layers. By combining the lower two leukocyte
layers (M2/3), a population of neutrophils consisting of 26% bands and 63%
PMNs is seen. When compared with peripheral blood PMNs, these bone marrow
neutrophils had a lower alkaline phosphatase activity, decreased ingestion
of Oil Red O-coated particles, impaired superoxide release on stimulation
with the chemotactic peptide Fmet-leu-phe (FMLP) or the tumor promotor
phorbol myristate acetate (PMA), and less activity of the NADPH-dependent
oxidase. These results indicate that morphologically mature neutrophilic
cells within the bone marrow exist in a still functionally immature state.
Volume 68,
Issue 4,
pp. 853-860,
10/01/1986
Copyright © 1986 by The American Society of Hematology
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