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Granulocyte-macrophage colony-stimulating factor (GM-CSF) in human long-
term bone marrow cultures: endogenous production in the adherent layer and
effect of exogenous GM-CSF on granulomonopoiesis
P Charbord, E Tamayo, S Saeland, V Duvert, J Poulet, AM Gown and P Herve
INSERM/Centre Regional de Transfusion Sanguine, Besancon, France.
This study was designed to assess the presence of endogenous
granulocyte-macrophage colony-stimulating factor (GM-CSF) within adherent
layers of human Dexter-type cultures and to investigate the effect on
granulomonopoiesis of adding exogenous GM-CSF to the culture medium. The
presence of GM-CSF was demonstrated using a bioassay, in which adherent
layers from normal bone marrows gave rise to endogenous
granulocyte-macrophage colony-forming units (CFU-GM) that were specifically
inhibited by increasing amounts of an anti-GM-CSF neutralizing antibody.
Using an immunoassay, the estimated amounts of GM-CSF were less than or
equal to 40 pg per flask in adherent layers, while remaining undetectable
in supernatants. The addition of 10 ng or purified recombinant GM-CSF per
milliliter of culture medium increased slightly the CFU-GM output over a
5-week culture period. The addition of 50 ng/mL decreased significantly the
CFU-GM output after 5 weeks of culture. This decrease was associated with
major modifications of the adherent layer cell composition. Large round or
ovoid macrophages were generated at the expense of the interdigitated and
elongated stromal cells and the extracellular fibronectin network was no
longer observed. These studies suggest that GM-CSF production by accessory
cells (stromal cells and/or monocytes) is almost equal to its consumption
by hematopoietic cells, a situation similar to that found in long-term
cultures of murine marrows. They also show that the maintenance of
granulomonopoiesis is decreased by adding more than 10 ng/mL of exogenous
GM-CSF to the culture medium, which is related to the induction of adherent
macrophages, the disappearance of the major smooth-muscle-like stromal cell
component of the adherent layer, and that of the fibronectin extracellular
matrix.
Volume 78,
Issue 5,
pp. 1230-1236,
09/01/1991
Copyright © 1991 by The American Society of Hematology
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