title: Gene expression profiles of human chondrocytes during passaged monolayer cultivation.
abstract: Chondrocyte phenotype has been shown to dedifferentiate during passaged monolayer cultivation. Hence, we have investigated the expression profile of 27 chondrocyte-associated genes from both osteoarthritic cartilage tissue and healthy passaged human articular chondrocytes by quantitative real-time PCR. Our results indicate that the gene expression levels of matrix proteins and proteases in chondrocytes from monolayer culture decrease compared with those from cartilage tissue, while monolayer cultured chondrocytes from normal and osteoarthritic cartilage exhibit similar gene expression patterns. However, chondrocytic gene expression profiles were differentially altered at various stages of passage. The expression of the matrix proteins aggrecan, type II collagen, and fibromodulin inversely correlated with increasing passage number, while fibronectin and link protein exhibited a marked increase with passage. The expression of matrix proteinases MMP-3/9/13 and ADAMTS-4/5 decreased with passage, whereas proteinase inhibitors TIMP-2/3 were elevated. The cytokine IL-1 also showed increased expression with monolayer chondrocyte culture, while IGF-1 expression levels were diminished. No significant changes in TGF-beta, or the chondrogenic transcription factors Sox-9, c-fos, or c-jun were observed. Our data indicates that cultured chondrocytes undergo dedifferentiation during monolayer culture, although the gene expression level of transcription factors necessary for chondrogenesis remains unchanged. This data may prove important for the future development of more specific and efficacious cultivation techniques for human articular chondrocyte-based therapies.
authors.affiliation: Centre of Orthopaedic Surgery, University of Western Australia, M Block, QEII Medical Centre, Monash Avenue, Nedlands, Western Australia 6009, Australia.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Lin
authors.forename: Jonathan B
authors.initials: JB
authors.lastname: Fitzgerald
authors.forename: Jiake
authors.initials: J
authors.lastname: Xu
authors.forename: Craig
authors.initials: C
authors.lastname: Willers
authors.forename: David
authors.initials: D
authors.lastname: Wood
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Grodzinsky
authors.forename: Ming H
authors.initials: MH
authors.lastname: Zheng
mesh_terms: D051722:ADAM Proteins / Q000235:genetics; D000071121:ADAMTS4 Protein; D000071116:ADAMTS5 Protein; D053674:Aggrecans / Q000235:genetics; D002358:Cartilage, Articular / Q000378:metabolism*; D018929:Cell Culture Techniques; D019902:Chondrocytes / Q000378:metabolism*; D024043:Collagen Type II / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics; D000071235:Fibromodulin; D005353:Fibronectins / Q000235:genetics; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics; D007375:Interleukin-1 / Q000235:genetics; D007719:Knee Joint / Q000378:metabolism*; D053505:Matrix Metalloproteinases, Secreted / Q000235:genetics; D016133:Polymerase Chain Reaction; D011348:Procollagen N-Endopeptidase / Q000235:genetics; D011509:Proteoglycans / Q000235:genetics; D012313:RNA / Q000302:isolation & purification; D048348:Reverse Transcription; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000235:genetics; D000099249:Proteoglycan Link Protein
chemical_list: D051722:ADAM Proteins; D000071121:ADAMTS4 Protein; D000071116:ADAMTS5 Protein; D053674:Aggrecans; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D000071235:Fibromodulin; D005353:Fibronectins; D007334:Insulin-Like Growth Factor I; D007375:Interleukin-1; D053505:Matrix Metalloproteinases, Secreted; D011348:Procollagen N-Endopeptidase; D011509:Proteoglycans; D012313:RNA; D019716:Tissue Inhibitor of Metalloproteinase-2; D019717:Tissue Inhibitor of Metalloproteinase-3; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D000099249:Proteoglycan Link Protein; C000604330:FMOD protein, human; C088390:TIMP3 protein, human; C121532:ADAMTS5 protein, human; C000603918:ADAMTS4 protein, human