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Chondrocyte cluster formation in agarose cultures as a functional assay to identify genes expressed in osteoarthritis.

Quintavalla, Joseph and Kumar, Chandrika and Daouti, Sherif and Slosberg, Eric and Uziel-Fusi, Susan (2005) Chondrocyte cluster formation in agarose cultures as a functional assay to identify genes expressed in osteoarthritis. Journal of Cellular Physiology, 204 (2). pp. 560-566. ISSN 0021-9541

Abstract

Understanding altered gene expression in osteoarthritic cartilage can lead to new targets for drug intervention. We established a functional assay based on chondrocyte cluster formation, a phenotype associated with osteoarthritis (OA), to screen an OA cartilage gene library. Previous reports have demonstrated that normal chondrocytes grown in suspension culture maintain their chondrocytic phenotype, however, certain growth factors such as basic fibroblast growth factor (bFGF) will induce the cells to proliferate in tight clusters similar to those seen in osteoarthritic cartilage. In this study we validate that overexpression of bFGF by retrovirally transduced normal chondrocytes would similarly induce the proliferation of tight cell clusters. We then used this approach as a basis to set up a functional screen where an entire OA cartilage cDNA library was tranduced into normal chondrocytes to search for other genes that would also induce cluster formation. Seven potential genes were isolated from the OA gene library, including BPOZ, IL-17 receptor C, NADH ubiquinone oxidoreductase, COMP, Soluble carrier 16 (MCT 3), C1r, and bFGF itself. None of the identified genes were upregulated by bFGF, however, all of them upregulated the expression of bFGF suggesting a common pathway. Although cluster formation is not considered to be destructive in OA cartilage, it is consistent with the disease and could yield answers to the altered phenotype. Further studies are needed to elucidate how these genes are linked to the disease state.

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Date Deposited: 14 Dec 2009 13:56
Last Modified: 14 Dec 2009 13:56
URI: https://oak.novartis.com/id/eprint/654

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