1α,25-Dihydroxyvitamin D3 [1,25(OH)2D3] is known to modulate Ca2+ metabolism in several cell types. Vitamin-D-dependent calcium binding proteins such as calbindin-D28K (28 kDa calcium binding proteins) have been shown to be regulated by 1,25(OH)2D3 but the mechanisms controlling calbindin synthesis are still poorly understood in human osteoblast cell culture models. The human bone marrow stromal cells (HBMSC) described in this paper developed a calcified matrix, expressed osteocalcin (OC), osteopontin (OP) and responded to 1,25(OH)2D3. The expression of vitamin D receptor mRNA was demonstrated by reverse transcription-PCR. Calbindin-D28K protein was identified only in cells arising from the sixth subculture, which exhibited a calcified matrix and all of the osteoblastic markers, e.g. OC and OP. It was demonstrated by dot-immunodetection using immunological probes, and by in situ hybridization using labelled cDNA probes. Moreover, vitamin D3 enhanced calbindin-D28K synthesis as well as OC synthesis and alkaline phosphatase activity. Uptake of 45Ca induced into the matrix by 1,25(OH)2D3 supports the hypothesis that the calcium-enriched matrix could trap calbindin-D proteins. In conclusion, the studies in vitro described in the present paper indicate, for the first time, a possible role of calbindin-D28K in mineralized matrix formation in HBMSC.
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August 1998
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Research Article|
August 01 1998
Synthesis of calbindin-D28K during mineralization in human bone marrow stromal cells
Corinne FAUCHEUX;
Corinne FAUCHEUX
1INSERM U 443, Université Victor Ségalen Bordeaux 2, 146 rue Léo-Saignat, 33076 Bordeaux cedex, France
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Reine BAREILLE;
Reine BAREILLE
1INSERM U 443, Université Victor Ségalen Bordeaux 2, 146 rue Léo-Saignat, 33076 Bordeaux cedex, France
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Joëlle AMEDEE
Joëlle AMEDEE
1
1INSERM U 443, Université Victor Ségalen Bordeaux 2, 146 rue Léo-Saignat, 33076 Bordeaux cedex, France
1To whom correspondence should be addressed (e-mail joelle.amedee@bordeaux.inserm.fr).
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Publisher: Portland Press Ltd
Received:
November 27 1997
Revision Received:
April 09 1998
Accepted:
May 08 1998
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 333 (3): 817–823.
Article history
Received:
November 27 1997
Revision Received:
April 09 1998
Accepted:
May 08 1998
Citation
Corinne FAUCHEUX, Reine BAREILLE, Joëlle AMEDEE; Synthesis of calbindin-D28K during mineralization in human bone marrow stromal cells. Biochem J 1 August 1998; 333 (3): 817–823. doi: https://doi.org/10.1042/bj3330817
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