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Changes in matrix protein gene expression associated with mineralization in the differentiating chick limb‐bud micromass culture system
Author(s) -
Teixeira Cristina C.,
Xiang Jenny,
Roy Rani,
Kudrashov Valery,
Binderman Itzhak,
MayerKuckuk Philipp,
Boskey Adele L.
Publication year - 2011
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.22951
Subject(s) - bone sialoprotein , osteopontin , limb bud , mineralization (soil science) , alkaline phosphatase , microbiology and biotechnology , gene expression , chemistry , dmp1 , matrix gla protein , biology , biochemistry , phosphate , gene , osteocalcin , endocrinology , enzyme , viral matrix protein , hyperphosphatemia , organic chemistry , nitrogen
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. J. Cell. Biochem. 112: 607–613, 2011. © 2010 Wiley‐Liss, Inc.