Biochemical and Biophysical Research Communications
Vitronectin and collagen I differentially regulate osteogenesis in mesenchymal stem cells
Section snippets
Materials and methods
Materials. Purified type-I collagen and human vitronectin were purchased from Cohesion (Palo Alto, CA) and Chemicon International (Temecula, CA), respectively. Phospho-p44/42 MAPK antibody (anti-phospho-ERK1/2, monoclonal) was purchased from Cell Signaling Technology, Inc. (Beverly, MA). Antibodies to ERK1, paxillin, and p-FAK (Y397) were purchased from BD Transduction Laboratories (Lexington, KY). Anti-pser473-Akt (rabbit polyclonal) and anti-pY31-paxillin (rabbit polyclonal) antibodies were
Integrin expression and utilization in MSCs
Based in part on our recent report that MSCs undergo osteogenesis to differing degrees when cultured on thin polymer films due to the differential deposition of serum-derived ECM proteins [32], we hypothesized that MSC differentiation must be influenced by integrins. Our first goal in testing this hypothesis was to use flow cytometry to quantify the integrin expression repertoire and adhesion blocking to determine which integrins are actually used to adhere to Col I and VN. Flow cytometric
Discussion
It is widely recognized that cell–ECM interactions play an important role in tissue development [37], [38], and that signaling pathways initiated by integrin binding are involved in regulating nearly every aspect of cell function, including cell cycle progression, migration, apoptosis, and differentiation [16], [39], [40]. In this study, we investigated the impact of integrin-mediated adhesion to vitronectin and type-I collagen on the osteogenic differentiation of MSCs. Our exclusive focus on
Acknowledgments
This work was partially supported by a grant from the NIH/NIDCR (1R03-DE016117) awarded to A.J.P. and by internal set-up funds provided by UC Irvine. In addition, we acknowledge Aswathi Sreedharan for cell culture assistance, Cyrus Ghajar for critically reading the manuscript, and the UC Irvine Center for Immunology for assistance with flow cytometry.
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