title: Human chondrocyte cultures as models of cartilage-specific gene regulation.
abstract: The human adult articular chondrocyte is a unique cell type that has reached a fully differentiated state as an end point of development. Within the cartilage matrix, chondrocytes are normally quiescent and maintain the matrix constituents in a low-turnover state of equilibrium. Isolated chondrocytes in culture have provided useful models to study cellular responses to alterations in the environment such as those occurring in different forms of arthritis. However, expansion of primary chondrocytes in monolayer culture results in the loss of phenotype, particularly if high cell density is not maintained. This chapter describes strategies for maintaining or restoring differentiated phenotype by culture in suspension, gels, or scaffolds. Techniques for assessing phenotype involving primarily the analysis of synthesis of cartilage-specific matrix proteins as well as the corresponding mRNAs are also described. Approaches for studying gene regulation, including transfection of promoter-driven reporter genes with expression vectors for transcriptional and signaling regulators, chromatin immunoprecipitation, and DNA methylation are also described.
authors: Laboratory for Cartilage Biology, The Hospital for Special Surgery, Weill Cornell Medical College, New York, NY, USA.
authors: Miguel
authors: M
authors: Otero
authors: Marta
authors: M
authors: Favero
authors: Cecilia
authors: C
authors: Dragomir
authors: Karim El
authors: KE
authors: Hachem
authors: Ko
authors: K
authors: Hashimoto
authors: Darren A
authors: DA
authors: Plumb
authors: Mary B
authors: MB
authors: Goldring
mesh_terms: D002356:Cartilage / Q000166:cytology / Q000378:metabolism*; D064236:Cartilage Oligomeric Matrix Protein; D002469:Cell Separation / Q000379:methods; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D019175:DNA Methylation; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D064235:Matrilin Proteins; D061251:Primary Cell Culture / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D064236:Cartilage Oligomeric Matrix Protein; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D064235:Matrilin Proteins; D012333:RNA, Messenger; C577553:TSP5 protein, human