title: Isolation and characterization of rheumatoid arthritis synovial fibroblasts from primary culture--primary culture cells markedly differ from fourth-passage cells.
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-1beta. 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.
authors.affiliation: Experimental Rheumatology Unit, Friedrich Schiller University Jena, Jena, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Zimmermann
authors.forename: E
authors.initials: E
authors.lastname: Kunisch
authors.forename: R
authors.initials: R
authors.lastname: Pfeiffer
authors.forename: A
authors.initials: A
authors.lastname: Hirth
authors.forename: H D
authors.initials: HD
authors.lastname: Stahl
authors.forename: U
authors.initials: U
authors.lastname: Sack
authors.forename: A
authors.initials: A
authors.lastname: Laube
authors.forename: E
authors.initials: E
authors.lastname: Liesaus
authors.forename: A
authors.initials: A
authors.lastname: Roth
authors.forename: E
authors.initials: E
authors.lastname: Palombo-Kinne
authors.forename: F
authors.initials: F
authors.lastname: Emmrich
authors.forename: R W
authors.initials: RW
authors.lastname: Kinne
mesh_terms: D001172:Arthritis, Rheumatoid / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D005347:Fibroblasts / Q000473:pathology*; D006801:Humans; D010003:Osteoarthritis / Q000473:pathology; D013583:Synovial Membrane / Q000473:pathology*