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Stromal cell progeny of murine bone marrow fibroblast colony-forming units
are clonal endothelial-like cells that express collagen IV and laminin
S Perkins and RA Fleischman
Department of Internal Medicine, University of Texas Southwestern Medical
Center, Dallas 75235.
Studies of human and murine bone marrow explants have demonstrated the
existence of stromal cell precursors that give rise to colonies of adherent
cells in short-term cultures. Because previous data suggested that these
colonies were composed of fibroblasts, the precursor cells were termed
fibroblast colony-forming units (CFU-F). However, we have recently shown
that the stromal cells which support hematopoiesis in murine long-term bone
marrow cultures (LTBC) express collagen IV and laminin, markers associated
with an endothelial cell lineage, but are negative for collagen I and III,
markers associated with a fibroblast cell lineage. Because these
conflicting results suggest major functional differences between the
stromal cells observed in long-term cultures and the short-term assay, we
re-examined the lineage of CFU-F- derived stromal cells. Using two-color
immunofluorescence, we characterized virtually all of the cells comprising
individual "CFU-F" colonies derived from mouse radiation chimeras.
Identification of donor (hematopoietic) or host (stromal) origin was based
on surface staining for strain-specific H-2 surface antigens, and, for
endothelial or fibroblast properties, on cytoplasmic staining for laminin
and collagen IV, or collagens I and III, respectively. The results
demonstrate that a large proportion of the cells in CFU-F colonies are
donor-derived and fail to stain with any of the antisera specific for
nonhematopoietic cells. In addition, these donor-derived cells exhibit
marked phagocytic capacity and stain positively with monoclonal antibodies
characteristic of the monocyte-macrophage hematopoietic cell lineage
(anti-T200, anti- Mac-1, F4/80). However, the remainder of the cells are
host-derived cells that stain positively with antisera to collagen IV and
laminin. In contrast, stains for collagen types I and III were negative
under conditions that allowed for strong staining of control skin
fibroblasts. In separate studies, using mixtures of two genetically
distinct bone marrows, the cells expressing collagen IV were further shown
to be clonal in origin within individual colonies, directly demonstrating
that the CFU-F assay provides a quantitative measure of the numbers of
marrow stromal cell precursors. Thus, the current studies establish a
remarkable similarity between the hematopoietic microenvironment in the
short-term CFU-F assay and the long-term culture system: the majority of
adherent cells are hematopoietic cells of the monocyte-macrophage lineage,
while the remainder are stromal cells whose precise lineage remains
uncertain, but whose pattern of collagen expression is more consistent with
an endothelial rather than a fibroblast cell origin.
Volume 75,
Issue 3,
pp. 620-625,
02/01/1990
Copyright © 1990 by The American Society of Hematology
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