Changes in matrix protein gene expression associated with mineralization in the differentiating chick limb-bud micromass culture system.
Chick limb-bud mesenchymal stem cells plated in high density culture in the presence of 4 mM inorganic phosphate and vitamin C differentiate and form a mineralizable matrix, resembling that of the chick growth plate. To further elucidate the mechanism that allows these cultures to form physiologic hydroxyapatite deposits, and how the process can be manipulated to gain insight into mineralization mechanisms, we compared gene expression in mineralizing (with 4 mM inorganic phosphate) and non-mineralizing cultures (containing only 1 mM inorganic phosphate) at the start of mineralization (day 11) and after mineralization reached a plateau (day 17) using a chick specific microarray. Based on replicate microarray experiments and K-cluster analysis, several genes associated with the mineralization process were identified, and their expression patterns confirmed throughout the culture period by quantitative RT-PCR. The functions of bone morphogenetic protein 1, BMP1, dentin matrix protein 1, DMP1, the sodium phosphate co-transporter, NaPi IIb, matrix metalloprotease 13. MMP-13, and alkaline phosphatase, along with matrix protein genes (type X collagen, bone sialoprotein, and osteopontin) usually associated with initiation of mineralization are discussed.
New York University, College of Dentistry, New York, New York, USA.
Cristina C
CC
Teixeira
Jenny
J
Xiang
Rani
R
Roy
Valery
V
Kudrashov
Itzhak
I
Binderman
Philipp
P
Mayer-Kuckuk
Adele L
AL
Boskey
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002642:Chick Embryo; D002645:Chickens; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D018878:Limb Buds / Q000166:cytology* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D016326:Extracellular Matrix Proteins
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