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Biologic significance of constitutive and subliminal growth factor
production by bone marrow stroma
EL Kittler, H McGrath, D Temeles, RB Crittenden, VK Kister and PJ Quesenberry
Department of Internal Medicine, University of Virginia Health Sciences
Center, Charlottesville 22908.
The "stromal" or adherent cells of long-term murine Dexter explant bone
marrow cultures provide the best in vitro model of the bone marrow
microenvironment. Colony-stimulating factor-1 (CSF-1) is produced
constitutively by these cells and is easily detected, but most
investigators have not found constitutive production of the other
hemolymphopoietic cytokines. We have previously reported the detection of
granulocyte-macrophage-CSF (GM-CSF) in murine stromal cultures and its
induction by the lectin Pokeweed mitogen. The present studies analyzing
stromal cytokine messenger RNA (mRNA) production by standard Northern blot
analysis show constitutive production of mRNAs for CSF-1, GM-CSF,
granulocyte-CSF (G-CSF), c-kit ligand (KL), and interleukin-6 (IL-6), but
not IL-3, IL-4, or IL-5 by 3-week irradiated or nonirradiated murine Dexter
stromal cells. Exposure of stromal cells to Pokeweed mitogen or IL-1 16
hours before RNA harvest induces the messages for GM-CSF, G-CSF, KL, and
IL-6, but not IL-3, IL-4, IL-5, or CSF-1. Polymerase chain reaction
amplification of cDNA made with reverse transcriptase from stromal RNA
using two separate sets of IL-3- specific primers shows the presence of
IL-3 message in irradiated stromal cells, which is only detectable with
this more sensitive technique. The factor-dependent cell lines FDC-P1 and
32D are supported by the stromal cells without the addition of exogenous
growth factors, demonstrating a cytokine activity in these cultures that is
inhibited by the addition of anti-IL-3 or anti-GM-CSF antibodies. These
data indicate that murine Dexter stromal cells constitutively produce
CSF-1, GM-CSF, G-CSF, IL-6, KL, and IL-3. This growth factor production
could explain the support of granulocyte, macrophage, and megakaryocyte
production and stem cell maintenance in Dexter-type long-term murine bone
marrow cultures.
Volume 79,
Issue 12,
pp. 3168-3178,
06/15/1992
Copyright © 1992 by The American Society of Hematology
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