Gene expression profile of human bone marrow stromal cells determined by restriction fragment differential display analysis

• Published in: Journal of cellular biochemistry Vol. 92; no. 4; pp. 733 - 744
• Main Authors: Monticone, Massimiliano, Liu, Yi, Tonachini, Laura, Mastrogiacomo, Maddalena, Parodi, Stefano, Quarto, Rodolfo, Cancedda, Ranieri, Castagnola, Patrizio
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.07.2004
• Subjects: Adolescent; Adult; Amino Acid Oxidoreductases - genetics; Amino Acid Oxidoreductases - metabolism; Bone Marrow Cells - metabolism; cancer; Cells, Cultured; differential display; European Continental Ancestry Group; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Gene Expression; gene expression profile; Gene Expression Profiling; human bone marrow stromal cells; Humans; IGFbp-3; Insulin-Like Growth Factor Binding Protein 3 - genetics; Insulin-Like Growth Factor Binding Protein 3 - metabolism; LOXL2; Middle Aged; Osteoblasts - metabolism; Polymerase Chain Reaction; Restriction Mapping; Stromal Cells - metabolism; Transforming Growth Factor beta - genetics; Transforming Growth Factor beta - metabolism; βIG-h3
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.20120

Using an in vitro osteogenic culture system, we carried out a restriction fragment differential display (RFDD‐PCR) to identify genes expressed by these cells in their undifferentiated stage and not expressed, or expressed at a lower level, in a closely related but distinct cell type: bone marrow stromal cells (BMSC)‐derived osteoblasts (BDO). Forty‐seven candidate regulated genes, selected by RFDD, were analyzed by RT‐PCR analysis in three cell clones and in primary cultures from seven different donors. A subset of three genes were confirmed as upregulated in BMSC relative to BDO in every primary culture and cloned population examined: βIG‐h3, IGFbp3, and LOXL2. Their differential expression was confirmed by Northern analysis and the corresponding proteins were detected by immunolocalization in BMSC. © 2004 Wiley‐Liss, Inc.