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Despite a growing use of FLS isolated from murine inflammatory models, a detailed characterisation of these cells has not been performed.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>In this study, FLS were isolated from inflamed joints of mice expressing both the T cell receptor transgene KRN and the MHC class II molecule Ag7 (K\/BxN mice) and their purity in culture determined by immunofluorescence and real-time reverse transcription polymerase chain reaction (real-time RT-PCR). Basal expression of proinflammatory genes was determined by real-time RT-PCR. Secreted interleukin 6 (IL-6) was measured by enzyme-linked immunosorbent assay (ELISA), and its regulation by tumor necrosis factor-alpha (TNF-\u03b1 and corticosterone (the major glucocorticoid in rodents) measured relative to other mesenchymal cell populations.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Purity of FLS culture was identified by positive expression of fibronectin, prolyl 4-hydroxylase, cluster of differentiation 90.2 (CD90.2) and 248 (CD248) in greater than 98% of the population. Cultured FLS were able to migrate and invade through matrigel, a process enhanced in the presence of TNF-\u03b1. FLS isolated from K\/BxN mice possessed significantly greater basal expression of the inflammatory markers IL-6, chemokine ligand 2 (CCL-2) and vascular cell adhesion molecule 1 (VCAM-1) when compared to FLS isolated from non-inflamed tissue (IL-6, 3.6 fold; CCL-2, 11.2 fold; VCAM-1, 9 fold;\n                      <jats:italic>P<\/jats:italic>\n                      &lt; 0.05). This elevated expression was abrogated in the presence of corticosterone at 100 nmol\/l. TNF-\u03b1 significantly increased expression of all inflammatory markers to a much greater degree in K\/BxN FLS relative to other mesenchymal cell lines (K\/BxN; IL-6, 40.8 fold; CCL-2, 1343.2 fold; VCAM-1, 17.8 fold; ICAM-1, 13.8 fold;\n                      <jats:italic>P<\/jats:italic>\n                      &lt; 0.05), with secreted IL-6 mirroring these results (K\/BxN; con, 169 \u00b1 29.7 versus TNF-\u03b1, 923 \u00b1 378.8 pg\/ml\/1 \u00d7 10\n                      <jats:sup>5<\/jats:sup>\n                      cells;\n                      <jats:italic>P<\/jats:italic>\n                      &lt; 0.05). Dose response experiments confirmed effective concentrations between 10 and 100 nmol\/l for corticosterone and 1 and 10 ng\/ml for TNF-\u03b1, whilst inflammatory gene expression in FLS was shown to be stable between passages four and seven.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      This study has established a well characterised set of key inflammatory genes for\n                      <jats:italic>in vitro<\/jats:italic>\n                      FLS culture, isolated from K\/BxN mice and non-inflamed wild-type controls. Their response to both pro- and anti-inflammatory signalling has been assessed and shown to strongly resemble that which is seen in human FLS culture. Additionally, this study provides guidelines for the effective characterisation, duration and treatment of murine FLS culture.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/ar4158","type":"journal-article","created":{"date-parts":[[2013,1,30]],"date-time":"2013-01-30T12:14:36Z","timestamp":1359548076000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":57,"title":["Characterisation of fibroblast-like synoviocytes from a murine model of joint inflammation"],"prefix":"10.1186","volume":"15","author":[{"given":"Rowan S","family":"Hardy","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"H\u00fclso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingling","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylvia J","family":"Gasparini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colette","family":"Fong-Yee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinwen","family":"Tu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shihani","family":"Stoner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul M","family":"Stewart","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karim","family":"Raza","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark S","family":"Cooper","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus J","family":"Seibel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2013,1,29]]},"reference":[{"key":"3896_CR1","doi-asserted-by":"publisher","first-page":"2619","DOI":"10.1002\/1529-0131(200012)43:12<2619::AID-ANR1>3.0.CO;2-V","volume":"43","author":"PP Tak","year":"2000","unstructured":"Tak PP, Bresnihan B: The pathogenesis and prevention of joint damage in rheumatoid arthritis: advances from synovial biopsy and tissue analysis. 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