title: Immortalized cell lines from mouse xiphisternum preserve chondrocyte phenotype.
abstract: Chondrocytes are unique to cartilage and the study of these cells in vitro is important for advancing our understanding of the role of these cells in normal homeostasis and disease including osteoarthritis (OA). As there are limitations to the culture of primary chondrocytes, cell lines have been developed to overcome some of these obstacles. In this study, we developed a procedure to immortalize and characterize chondrocyte cell lines from mouse xiphisternum. The cells displayed a polygonal to fibroblastic morphology in monolayer culture. Gene expression studies using quantitative PCR showed that the cell lines responded to bone morphogenetic protein 2 (BMP-2) by increased expression of matrix molecules, aggrecan, and type II collagen together with transcriptional factor, Sox9. Stimulation by IL-1 results in the increased expression of catabolic effectors including MMP-13, nitric oxide synthase, ADAMTS4, and ADAMTS5. Cells cultured in alginate responded to BMP-2 by increased synthesis of proteoglycan (PG), a major matrix molecule of cartilage. IL-1 treatment of cells in alginate results in increased release of PG into the conditioned media. Further analysis of the media showed the presence of Aggrecanase-cleaved aggrecan fragments, a signature of matrix degradation. These results show that the xiphisternum chondrocyte cell lines preserve their chondrocyte phenotype cultured in either monolayer or 3-dimensional alginate bead culture systems. In summary, this study describes the establishment of chondrocyte cell lines from the mouse xiphisternum that may be useful as a surrogate model system to understand chondrocyte biology and to shed light on the underlying mechanism of pathogenesis in OA.
authors.affiliation: Wyeth Research, Department of Women's Health and Musculoskeletal Biology, Cambridge, Massachusetts 02140, USA. mmajumdar@wyeth.com
authors.forename: Manas K
authors.initials: MK
authors.lastname: Majumdar
authors.forename: Priya S
authors.initials: PS
authors.lastname: Chockalingam
authors.forename: Ramesh A
authors.initials: RA
authors.lastname: Bhat
authors.forename: Richard
authors.initials: R
authors.lastname: Sheldon
authors.forename: Cristin
authors.initials: C
authors.lastname: Keohan
authors.forename: Tracey
authors.initials: T
authors.lastname: Blanchet
authors.forename: Sonya
authors.initials: S
authors.lastname: Glasson
authors.forename: Elisabeth A
authors.initials: EA
authors.lastname: Morris
mesh_terms: D053674:Aggrecans; D000464:Alginates / Q000378:metabolism; D000818:Animals; D015415:Biomarkers; D002356:Cartilage / Q000166:cytology; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D007375:Interleukin-1 / Q000494:pharmacology; D037181:Lectins, C-Type / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009569:Nitric Oxide / Q000096:biosynthesis; D010641:Phenotype; D011509:Proteoglycans / Q000378:metabolism; D013249:Sternum / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
chemical_list: C508320:Acan protein, mouse; D053674:Aggrecans; D000464:Alginates; D015415:Biomarkers; D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D006603:Hexuronic Acids; D007375:Interleukin-1; D037181:Lectins, C-Type; D011509:Proteoglycans; D009569:Nitric Oxide; D020723:Glucuronic Acid; D010450:Endopeptidases; C083641:aggrecanase