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

2.50
Hdl Handle:
http://hdl.handle.net/10541/74835
Title:
Retroviral transduction with SOX9 enhances re-expression of the chondrocyte phenotype in passaged osteoarthritic human articular chondrocytes.
Authors:
Tew, Simon R; Li, Ying; Pothacharoen, Peraphan; Tweats, Lisa M; Hawkins, Robert E; Hardingham, Timothy E
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.
Affiliation:
UK Centre for Tissue Engineering at The Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, UK.
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
Issue Date:
Jan-2005
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
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorTew, Simon R-
dc.contributor.authorLi, Ying-
dc.contributor.authorPothacharoen, Peraphan-
dc.contributor.authorTweats, Lisa M-
dc.contributor.authorHawkins, Robert E-
dc.contributor.authorHardingham, Timothy E-
dc.date.accessioned2009-07-21T16:55:36Z-
dc.date.available2009-07-21T16:55:36Z-
dc.date.issued2005-01-
dc.identifier.citationRetroviral transduction with SOX9 enhances re-expression of the chondrocyte phenotype in passaged osteoarthritic human articular chondrocytes. 2005, 13 (1):80-9 Osteoarthr. Cartil.en
dc.identifier.issn1063-4584-
dc.identifier.pmid15639641-
dc.identifier.doi10.1016/j.joca.2004.10.011-
dc.identifier.urihttp://hdl.handle.net/10541/74835-
dc.description.abstractOBJECTIVES: 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.en
dc.language.isoenen
dc.subject.meshCartilage, Articular-
dc.subject.meshCell Culture Techniques-
dc.subject.meshChondrocytes-
dc.subject.meshCollagen Type II-
dc.subject.meshExtracellular Matrix-
dc.subject.meshFibroblasts-
dc.subject.meshGene Expression Regulation-
dc.subject.meshGenetic Vectors-
dc.subject.meshGlycosaminoglycans-
dc.subject.meshHigh Mobility Group Proteins-
dc.subject.meshHumans-
dc.subject.meshInsulin-Like Growth Factor I-
dc.subject.meshOsteoarthritis, Knee-
dc.subject.meshPhenotype-
dc.subject.meshRetroviridae-
dc.subject.meshSOX9 Transcription Factor-
dc.subject.meshSkin-
dc.subject.meshTranscription Factors-
dc.subject.meshTransduction, Genetic-
dc.subject.meshTransforming Growth Factor beta-
dc.subject.meshTransforming Growth Factor beta3-
dc.titleRetroviral transduction with SOX9 enhances re-expression of the chondrocyte phenotype in passaged osteoarthritic human articular chondrocytes.en
dc.typeArticleen
dc.contributor.departmentUK Centre for Tissue Engineering at The Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, UK.en
dc.identifier.journalOsteoarthritis and Cartilageen
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