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Open Access Research article

Isolation and characterization of rheumatoid arthritis synovial fibroblasts from primary culture — primary culture cells markedly differ from fourth-passage cells

Thomas Zimmermann1, Elke Kunisch1, Robert Pfeiffer2, Astrid Hirth2, Hans-Detlev Stahl2, Ulrich Sack2, Anke Laube3, Eckehard Liesaus3, Andreas Roth3, Ernesta Palombo-Kinne1, Frank Emmrich2 and Raimund W Kinne1

Author affiliations

1 Experimental Rheumatology Unit, Friedrich Schiller University Jena, Jena, Germany

2 Institute of Clinical Immunology and Transfusion Medicine, University of Leipzig, Leipzig, Germany

3 Clinic of Orthopedics, Friedrich Schiller University Jena, Jena, Germany

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Citation and License

Arthritis Res 2001, 3:72-76  doi:10.1186/ar142

Published: 21 November 2000

Abstract

To reduce culture artifacts by conventional repeated passaging and long-term culture in vitro, the isolation of synovial fibroblasts (SFB) was attempted from rheumatoid arthritis (RA) synovial membranes by trypsin/collagenase digest, short-term in vitro adherence (7 days), and negative isolation using magnetobead-coupled anti-CD14 monoclonal antibodies. This method yielded highly enriched SFB (85% prolyl-4-hydroxylase+/74% Thy-1/CD90+ cells; <2% contaminating macrophages; <1% leukocytes/endothelial cells) that, in comparison with conventional fourth-passage RA-SFB, showed a markedly different phenotype and significantly lower proliferation rates upon stimulation with platelet-derived growth factor and IL-1β. This isolation method is simple and reliable, and may yield cells with features closer to the in vivo configuration of RA-SFB by avoiding extended in vitro culture.

Keywords:
isolation; phenotype/function; primary culture; rheumatoid arthritis; synovial fibroblasts