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    Retroviral transduction with SOX9 enhances re-expression of the chondrocyte phenotype in passaged osteoarthritic human articular chondrocytes.

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    Authors
    Tew, Simon R
    Li, Ying
    Pothacharoen, Peraphan
    Tweats, Lisa M
    Hawkins, Robert E
    Hardingham, Timothy E
    Affiliation
    UK Centre for Tissue Engineering at The Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, UK.
    Issue Date
    2005-01
    
    Metadata
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    Abstract
    OBJECTIVES: Articular chondrocytes proliferate in monolayer culture, but the expression of the transcription factor SOX9 falls and the ability of the cells to reform cartilage tissue declines. We have investigated whether retroviral SOX9 expression in extensively passaged human articular chondrocytes from osteoarthritic (OA) joints enables the cells to regain a cartilage matrix forming phenotype in pellet culture. DESIGN: Chondrocytes from normal and OA joints were retrovirally transduced with SOX9 and grown to passages 7-10 before being cultured as pellets of 500,000 cells for 14 days. Pellets were analysed by real time polymerase chain reaction, histology, immunohistochemistry and 1,9-dimethylmethylene blue assay. RESULTS: Chondrocytes from OA joints displayed higher expression of COL2A1 gene when transduced with SOX9 and cultured as pellets with 10% serum, but glycosaminoglycan (GAG) synthesis was low. Addition of transforming growth factor beta-3 and insulin like growth factor-1 increased collagen II expression and GAG synthesis in these SOX9 transduced cell pellets. The cells adopted a rounded morphology and there was increased deposition of collagen II protein compared to control green fluorescent protein transduced cell pellets. Similar results were seen with transduced chondrocytes from OA or healthy cartilage. SOX9 transduced human dermal fibroblasts did not show any chondrogenic response. DISCUSSION: Transduction with SOX9 primed the passaged articular chondrocytes to regain a chondrocytic phenotype in pellet culture and to form a cartilaginous matrix, which was enhanced by growth factors. Following transduction, chondrocytes from OA joints showed a similar capacity for chondrogenic recovery as those from healthy joints, which suggested that OA does not permanently compromise the chondrocyte phenotype.
    Citation
    Retroviral transduction with SOX9 enhances re-expression of the chondrocyte phenotype in passaged osteoarthritic human articular chondrocytes. 2005, 13 (1):80-9 Osteoarthr. Cartil.
    Journal
    Osteoarthritis and Cartilage
    URI
    http://hdl.handle.net/10541/74835
    DOI
    10.1016/j.joca.2004.10.011
    PubMed ID
    15639641
    Type
    Article
    Language
    en
    ISSN
    1063-4584
    ae974a485f413a2113503eed53cd6c53
    10.1016/j.joca.2004.10.011
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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