Gene expression profile of human bone marrow stromal cells determined by restriction fragment differential display analysis.
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: betaIG-h3, IGFbp3, and LOXL2. Their differential expression was confirmed by Northern analysis and the corresponding proteins were detected by immunolocalization in BMSC.
Monticone
Massimiliano
M
Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy. monticone@cba.unige.it
Liu
Yi
Y
Tonachini
Laura
L
Mastrogiacomo
Maddalena
M
Parodi
Stefano
S
Quarto
Rodolfo
R
Cancedda
Ranieri
R
Castagnola
Patrizio
P
D000293:Adolescent; D000328:Adult; D000594:Amino Acid Oxidoreductases / Q000235:genetics / Q000378:metabolism; D001854:Bone Marrow Cells / Q000378:metabolism*; D002478:Cells, Cultured; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D010006:Osteoblasts / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D017154:Stromal Cells / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D044465:White People; D000099222:betaIG-H3 Protein
D000594:Amino Acid Oxidoreductases; D016326:Extracellular Matrix Proteins; D018972:Insulin-Like Growth Factor Binding Protein 3; D016212:Transforming Growth Factor beta; D000099222:betaIG-H3 Protein; C119091:LOXL2 protein, human