Changes in matrix protein gene expression associated with mineralization in the differentiating chick limb-bud micromass culture system

Cristina C. Teixeira, Jenny Xiang, Rani Roy, Valery Kudrashov, Itzhak Binderman, Philipp Mayer-Kuckuk, Adele L. Boskey

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)607-613
Number of pages7
JournalJournal of Cellular Biochemistry
Volume112
Issue number2
DOIs
StatePublished - Feb 2011

Keywords

  • chick limb-bud
  • gene expression
  • microarray
  • micromass culture
  • mineralization

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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