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Fibroblast-like Synovial Cells Derived From Synovial Fluid JUDITH A. STEBULIS, RONALD G. ROSSETTI, FRANCISCO J. ATEZ, and ROBERT B. ZURIER
ABSTRACT. Methods. SF aspirated from joints of patients with rheumatoid arthritis (RA), other types of inflammatory arthritis, and osteoarthritis (OA) was centrifuged and the resulting cell pellet resuspended in growth medium. After 2 days, nonadherent cells were removed. FLS were also cultured from surgical specimens of synovial tissue (td-FLS). Phenotype characterization of fluid derived FLS (fd-FLS) was accomplished by flow cytometry and immunohistochemistry staining. Tumor necrosis factor-a (TNF-a) induced interleukin 6 (IL-6), IL-8, and cyclooxygenase 2 (COX-2) mRNA levels were assessed. Results. Second and later passage fd-FLS exhibited uniform fibroblast-like morphology. Fd-FLS and td-FLS expressed a similar profile of cell surface antigens including the fibroblast marker Thy-1. Less than 2% of either cell type expressed surface markers characteristic of dendritic cells, phagocytic cells, T cells, or leukocytes. Immunohistochemistry staining revealed the presence of fibroblast products prolyl-4 hydroxylase, procollagen I, and procollagen III in both culture types. TNF-a induced increases in IL-6, IL-8, and COX-2 mRNA were suppressed by dexamethasone in both fd-FLS and td-FLS. Conclusion. FLS can be cultured from SF. The fibroblast phenotype was confirmed by analysis of surface antigens and intracellular proteins. Inflammatory mediators produced after stimulation of both fd-FLS and td-FLS were suppressed by dexamethasone. In addition to providing a more accessible source of FLS, fd-FLS may also facilitate study of synovial cells in early RA when tissue specimens are not readily available. (J Rheumatol 2005;32:301-6) Key Indexing Terms:
SYNOVIAL CELLS
From the Department of Medicine, Rheumatology Division, University of Massachusetts Medical School, Worcester, Massachusetts, USA. Supported by NIH grants R01 AR3850 and T32 AR07572 from the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) (J. Stebulis, Trainee), R21 AT001471 from the National Center for Complementary and Alternative Medicine (NCCAM), R01 DA13691 from the National Institute on Drug Abuse (NIDA), and an Amgen Rheumatology Fellowship award to Dr. Stebulis. Core resources supported by the Diabetes Endocrinology Research Center grant DK32520 were also used. J.A. Stebulis, MD, Fellow; R.G. Rossetti, MPH, Research Associate; F.J. Atez, BS, Medical Student; R.B. Zurier, MD, Professor of Medicine. Address reprint requests to Dr. J.A. Stebulis, Department of Medicine, Rheumatology Division, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655. E-mail: judith.stebulis@umassmed.edu Submitted June 7, 2004; revision accepted November 9, 2004. |