Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro.

Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. Research Abstract Details 

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Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. Abstract Text:

Oliver Frank,Manuel Heim,Marcel Jakob,Andrea Barbero,Dirk ,Igor Bendik,Walter Dick,Michael Heberer,Ivan Martin,

We developed and used real-time RT-PCR assays to investigate how the expression of typical osteoblast-related genes by human bone marrow stromal cells (BMSC) is regulated by (i) the culture time in medium inducing osteogenic differentiation and (ii) the previous expansion in medium enhancing cell osteogenic commitment. BMSC from six healthy donors were expanded in medium without (CTR) or with fibroblast growth factor-2 and dexamethasone (FGF/Dex; these factors are known to increase BMSC osteogenic commitment) and further cultivated for up to 20 days with ascorbic acid, beta-glycerophosphate and dexamethasone (these factors are typically used to induce BMSC osteogenic differentiation). Despite a high variability in the gene expression levels among different individuals, we identified the following statistically significant patterns. The mRNA levels of bone morphogenetic protein-2 (BMP-2), bone sialo protein-II (BSP), osteopontin (OP) and to a lower extent cbfa-1 increased with culture time in osteogenic medium (OM), both in CTR- and FGF/Dex-expanded BMSC, unlike levels of alkaline phosphatase, collagen type I, osteocalcin, and osteonectin. After 20 days culture in OM, BMP-2, BSP, and OP were more expressed in FGF/Dex than in CTR-expanded BMSC (mRNA levels were, respectively, 9.5-, 14.9-, and 5.8-fold higher), unlike all the other investigated genes. Analysis of single-colony-derived strains of BMSC further revealed that after 20 days culture in OM, only a subset of FGF/Dex-expanded clones expressed higher mRNA levels of BMP-2, BSP, and OP than CTR-expanded clones. In conclusion, we provide evidence that mRNA levels of BMP-2, BSP, and OP, quantified using real-time RT-PCR, can be used as markers to monitor the extent of BMSC osteogenic differentiation in vitro; using those markers, we further demonstrated that only a few subpopulations of BMSC display enhanced osteogenic differentiation following FGF/Dex expansion.

Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. Publishing Authors By Initials

O Frank,M Heim,M Jakob,A Barbero,D ,I Bendik,W Dick,M Heberer,I Martin,

For similar peptides: intercellular signaling peptides and proteins: cytokines: transforming growth factor beta research abstracts see: peptides: intercellular signaling peptides and proteins: cytokines: transforming growth factor beta research

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Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of cellular biochemistry

VOLUME: 85

Page Numbers: 737-46

Journal Abbreviation: J. Cell. Biochem.

ISSN: 0730-2312

DAY: 19

MONTH: 11

YEAR: 2002

Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. Information

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LANGUAGE: eng

NlmUniqueID: 8205768

Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. Keywords Mesh Terms:

KEYWORDS: Transforming Growth Factor beta

MESH TERMS: genetics

Chemical & Substance for Abstract: Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. Information

Substance Name: Dexamethasone

Registry Number: 50-02-2

Grant and Affiliation Information for Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro.

AFFILIATION: Department of Surgery, Research Division, University of Basel, Switzerland.

Country: United States

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MEDLINETA: J Cell Biochem

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