Fibroblast-like synovial cells derived from synovial fluid.
Fibroblast-like synovial cells derived from synovial fluid.
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发表时间：
2005-02
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通讯作者：
Judith A. Stebulis;R. Rossetti;Francisco J Atez;R. Zurier
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作者：
Judith A. Stebulis;R. Rossetti;Francisco J Atez;R. Zurier
目的从滑液（SF）中获得成纤维细胞样滑膜细胞（FLS）。方法从类风湿关节炎（RA）、其他类型炎症性关节炎和骨关节炎（OA）患者的关节中抽吸SF，离心并将所得细胞颗粒重悬于生长培养基中。2天后，去除非贴壁细胞。从滑膜组织手术标本（td-FLS）中培养FLS。通过流式细胞术和免疫组织化学染色完成液体源性FLS （fd-FLS）的表型表征。评估肿瘤坏死因子- α (tnf - α)诱导的白细胞介素6 （IL-6）、IL-8和环氧化酶2 (COX-2) mRNA水平。结果二次传代及后续传代fd-FLS表现出均匀的成纤维细胞样形态。Fd-FLS和td-FLS表达类似的细胞表面抗原，包括成纤维细胞标记物Thy-1。两种细胞类型中表达树突状细胞、吞噬细胞、T细胞或白细胞特征的表面标记物均少于2%。免疫组织化学染色显示两种培养类型中均存在成纤维细胞产物脯氨酸-4羟化酶、前胶原I和前胶原III。TNF-a诱导的IL-6、IL-8和COX-2 mRNA升高在fd-FLS和td-FLS中均被地塞米松抑制。结论SF可培养FLS。通过表面抗原和细胞内蛋白分析证实成纤维细胞表型。地塞米松抑制fd-FLS和td-FLS刺激后产生的炎症介质。除了提供更容易获得的FLS来源外，fd-FLS还可以在无法获得组织标本的情况下促进早期RA滑膜细胞的研究。
OBJECTIVE To obtain fibroblast-like synovial cells (FLS) from synovial fluid (SF). 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-alpha (TNF-alpha) 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.