title: Immortalized mouse articular cartilage cell lines retain chondrocyte phenotype and respond to both anabolic factor BMP-2 and pro-inflammatory factor IL-1.
abstract: Articular cartilage chondrocytes help in the maintenance of tissue homeostasis and function of the articular joint. Study of primary chondrocytes in culture provides information closely related to in vivo functions of these cells. Limitations in the primary culture of chondrocytes have lead to the development of cells lines that serve as good surrogate models for the study of chondrocyte biology. In this study, we report the establishment and characterization of chondrocyte cell lines, MM-Sv/HP and MM-Sv/HP-2 from mouse articular cartilage. Cells were isolated from mouse femoral head articular cartilage, immortalized and maintained in culture through numerous passages. The morphology of the cells was from fibroblastic to polygonal in nature. Gene expression studies using quantitative PCR (Q-PCR) were performed on cells in monolayer culture and cells embedded in a three-dimensional alginate matrix. Stimulation of cells in monolayer culture with anabolic factor, BMP-2, resulted in increased gene expression of the extracellular matrix molecules, aggrecan and type II collagen and their regulator transcription factor, Sox9. Treatment by pro-inflammatory IL-1 resulted in increased gene expression of catabolic effectors including Aggrecanases (ADAMTS4, ADAMTS5), MMP-13 and nitric oxide synthase (Nos2). Cells in alginate treated with BMP-2 resulted in increased synthesis of proteoglycan which was released into the conditioned media on IL-1 stimulation. Western analysis of conditioned media showed the presence of Aggrecanase-cleaved aggrecan fragments. In summary, MM-Sv/HP and MM-Sv/HP-2 show preservation of important characteristics of articular chondrocytes as examined under multiple culture conditions and would provide a useful reagent in the study of chondrocyte biology.
authors.affiliation: Women's Health and Musculoskeletal Biology, Wyeth Research, Cambridge, Massachusetts, USA. manas.k.majumdar@gsk.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 / Q000378:metabolism; D000464:Alginates / Q000494:pharmacology; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002358:Cartilage, Articular / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D002461:Cell Line, Transformed; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D010450:Endopeptidases / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005269:Femur / Q000166:cytology / Q000187:drug effects; D006801:Humans; D018836:Inflammation Mediators / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology*; D051379:Mice; D010641:Phenotype; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
chemical_list: D053674:Aggrecans; D000464:Alginates; C525718:BMP2 protein, human; D015415:Biomarkers; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D018836:Inflammation Mediators; D007375:Interleukin-1; D016212:Transforming Growth Factor beta; D010450:Endopeptidases; C083641:aggrecanase