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Characterization of a 5-fluorouracil-enriched osteoprogenitor population of
the murine bone marrow
N Falla, Vlasselaer Van, J Bierkens, B Borremans, G Schoeters and U Van Gorp
Vlaamse Instelling voor Technologisch Onderzoek (VITO), Department of
Environment, Boeretang Mol Belgium, Geel.
In the presence of beta-glycerophosphate and vitamin C, cultures of normal
mouse bone marrow cells form three-dimensional structures that stain
positive with the Von Kossa technique and express alkaline phosphatase
(ALP), collagen type I, and osteocalcin. Little is known about the
characteristics and frequency of the cells that contribute to this
phenomenon. Most likely, mature osteoblastic cells do not contribute to the
nodule formation because no osteocalcin expressing cells are detected in
the flushed marrow by in situ hybridization. Limiting dilution analysis
shows that, in normal bone marrow, 1 of 2.2 x 10(5) cells has the potency
to form a bone nodule and to express ALP, collagen, and osteocalcin in a
temporal fashion. Upon in vivo treatment with 5-fluorouracil (5-FU), this
frequency increases 12-fold, eg, 1 in 1.75 x 10(4) cells shows osteogenic
activity. In comparison, fibroblast colony forming cells occur at a
frequency of 1 of 2.5 x 10(4) or 1 of 5 x 10(3) plated cells in normal or
5-FU-treated marrow, respectively. Using density centrifugation, the
majority of the osteoprogenitor cells in 5-FU marrow are found in the
low-density (1.066 to 1.067 g/mL) fractions. In addition, these cells bind
to nylon wool but not to plastic and aggregate in the presence of wheat
germ agglutinin and soybean agglutinin. Scanning and transmission electron
microscopy shows that the bone nodules in 5-FU marrow cultures are composed
of fibroblastoid cells embedded in a mineralized collagen matrix. In
conclusion, our results show that a quiescent cell population in the murine
bone marrow with fibroblastoid characteristics contributes to the formation
of bone-like nodules in vitro.
Volume 82,
Issue 12,
pp. 3580-3591,
12/15/1993
Copyright © 1993 by The American Society of Hematology
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