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Osteogenic progenitor cells in rat bone marrow stromal populations exhibit
self-renewal in culture
CA McCulloch, M Strugurescu, F Hughes, AH Melcher and JE Aubin
Faculty of Dentistry, University of Toronto, Ontario, Canada.
Marrow stromal cells are a heterogeneous population, comprising a variety
of lineages including osteogenic cells. In the presence of ascorbic acid,
sodium beta-glycerophosphate, and dexamethasone, rat bone marrow stromal
cells form discrete nodules of mineralized, bonelike tissue. We used nodule
formation by rat bone marrow stromal cells to assay for the self-renewal
capacity of osteogenic progenitor cell populations. Cultures were
subcultured every 5 days up to six times. Osteogenesis was assayed from
second to sixth subcultures by counting the number and measuring the areas
of mineralized nodules formed in cultures grown with 10(-8) mol/L
dexamethasone. Nodule number and area decreased progressively between
second and sixth subcultures. Alkaline phosphatase activity associated with
individual cells and measured videodensitometrically decreased
exponentially between the second and sixth subculture. The number of cells
with alkaline phosphatase activity also decreased with progressive
subculturing. The proportions of 3H-thymidine-labeled cells after
continuous labeling from the beginning of the culture period showed 90%
labeling for cells with alkaline phosphatase activity and fibroblastlike
cells. Cultures labeled for only the first 3 days exhibited higher labeling
of alkaline phosphatase-positive cells than fibroblastlike cells (P less
than .05). Cultures that were flash-labeled at the end of the culture
period demonstrated low labeling indices for cells with alkaline
phosphatase activity and up to 10-fold higher labeling indices for
fibroblastlike cells. Separate cultures treated with a cytocidal dose of
high specific activity 3H-thymidine did not form nodules. These results
indicate that osteogenic progenitor cells or another cell type required for
nodules to develop must divide early in culture if nodule formation is to
occur, and that osteoprogenitor cells express a limited capacity for
self-renewal.
Volume 77,
Issue 9,
pp. 1906-1911,
05/01/1991
Copyright © 1991 by The American Society of Hematology
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