TY - JOUR
T1 - Changes in matrix protein gene expression associated with mineralization in the differentiating chick limb-bud micromass culture system
AU - Teixeira, Cristina C.
AU - Xiang, Jenny
AU - Roy, Rani
AU - Kudrashov, Valery
AU - Binderman, Itzhak
AU - Mayer-Kuckuk, Philipp
AU - Boskey, Adele L.
PY - 2011/2
Y1 - 2011/2
N2 - Chick limb-bud mesenchymal stem cells plated in high density culture in the presence of 4mM 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 4mM inorganic phosphate) and non-mineralizing cultures (containing only 1mM 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.
AB - Chick limb-bud mesenchymal stem cells plated in high density culture in the presence of 4mM 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 4mM inorganic phosphate) and non-mineralizing cultures (containing only 1mM 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.
KW - chick limb-bud
KW - gene expression
KW - microarray
KW - micromass culture
KW - mineralization
UR - http://www.scopus.com/inward/record.url?scp=79251622447&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79251622447&partnerID=8YFLogxK
U2 - 10.1002/jcb.22951
DO - 10.1002/jcb.22951
M3 - Article
C2 - 21268082
AN - SCOPUS:79251622447
SN - 0730-2312
VL - 112
SP - 607
EP - 613
JO - Journal of Cellular Biochemistry
JF - Journal of Cellular Biochemistry
IS - 2
ER -