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The STRO-1+ fraction of adult human bone marrow contains the osteogenic
precursors
S Gronthos, SE Graves, S Ohta and PJ Simmons
Matthew Roberts Laboratory, Hanson Centre for Cancer Research, IMVS,
Adelaide, South Australia.
The monoclonal antibody STRO-1 identifies clonogenic bone marrow stromal
cell progenitors (fibroblast colony-forming units [CFU-F]) in adult human
bone marrow. These STRO-1+ CFU-F have previously been shown to give rise to
cells with the phenotype of fibroblasts, adipocytes, and smooth muscle
cells. In this study, the osteogenic potential of CFU- F derived from the
STRO-1+ fraction of adult human bone marrow was determined. CFU-F were
isolated from normal bone marrow aspirates by fluorescence activated cell
sorting, based on their expression of the STRO-1 antigen. Osteogenic
differentiation was assessed by the induction of alkaline phosphatase
expression, by the formation of a mineralized matrix (hydroxyapatite), and
by the production of the bone- specific protein osteocalcin. STRO-1+ cells
were cultured in the presence of dexamethasone (DEX; 10(-8) mol/L),
ascorbic acid 2- phosphate (ASC-2P; 100 mumol/L), and inorganic phosphate
(PO4i; 2.9 mmol/L). After 2 weeks of culture, greater than 90% of the cells
in each CFU-F colony stained positive for alkaline phosphatase using a
monoclonal antibody specific for bone and liver alkaline phosphatase.
Alkaline phosphatase activity was confirmed by histochemistry. A
mineralized matrix developed in the CFU-F cultures, after 4 weeks of
culture in the presence of DEX, ASC-2P, and PO4i. Mineralization was
confirmed by both light and electron microscopy. The mineral was identified
as hydroxyapatite by electron dispersive x-ray microanalysis and by x-ray
diffraction analysis. In replicate cultures, osteocalcin release was shown
after exposure of the cells to 1,25-dihydroxyvitamin D3 (10(-7) mol/L) both
by radioimmunoassay and Northern blot analysis. This work provides direct
evidence that adult human bone marrow-derived CFU-F are capable of
differentiating into functional osteoblasts and that osteoprogenitors are
present in the STRO-1+ population.
Volume 84,
Issue 12,
pp. 4164-4173,
12/15/1994
Copyright © 1994 by The American Society of Hematology
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