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Osteoblastic phenotype of rat marrow stromal cells cultured in the presence of dexamethasone, beta-glycerolphosphate, and L-ascorbic acidWe investigated the effects of the time course of addition of osteogenic supplements dexamethasone, beta-glycerolphosphate, and L-ascorbic acid to rat marrow stromal cells, and the exposure time on the proliferation and differentiation of the cells. It was the goal of these experiments to determine the time point for supplement addition to optimize marrow stromal cell proliferation and osteoblastic differentiation. To determine this, two studies were performed; one study was based on the age of the cells from harvest, and the other study was based on the duration of exposure to supplemented medium. Cells were seen to proliferate rapidly at early time points in the presence and absence of osteogenic supplements as determined by 3H-thymidine incorporation into the DNA of replicating cells. These results were supported by cell counts ascertained through total DNA analysis. Alkaline phosphatase (ALP) activity and osteocalcin production at 21 days were highest for both experimental designs when the cells were exposed to supplemented medium immediately upon harvest. The ALP levels at 21 days were six times greater for cells maintained in supplements throughout than for control cells cultured in the absence of supplements for both studies, reaching an absolute value of 75 x 10(-7) micromole/min/cell. Osteocalcin production reached 20 x 10(-6) ng/cell at 21 days in both studies for cells maintained in supplemented medium throughout the study, whereas the control cells produced an insignificant amount of osteocalcin. These results suggest that the addition of osteogenic supplements to marrow-derived cells early in the culture period did not inhibit proliferation and greatly enhanced the osteoblastic phenotype of cells in a rat model.
Document ID
20040172571
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Peter, S. J.
(Institute of Biosciences and Bioengineering, Rice University and Department of Chemical Engineering, Houston, Texas 77005-1892, United States)
Liang, C. R.
Kim, D. J.
Widmer, M. S.
Mikos, A. G.
McIntire, L. V.
Date Acquired
August 22, 2013
Publication Date
October 1, 1998
Publication Information
Publication: Journal of cellular biochemistry
Volume: 71
Issue: 1
ISSN: 0730-2312
Subject Category
Aerospace Medicine
Funding Number(s)
CONTRACT_GRANT: R01-AR44381
CONTRACT_GRANT: R29-AR41639
CONTRACT_GRANT: 5T32GM08362
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Cell Biology

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