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




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Arthritis Research and Therapy

, 3:72

First Online: 21 November 2000Received: 23 May 2000Revised: 10 October 2000Accepted: 17 October 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.

Keywordsisolation phenotype-function primary culture rheumatoid arthritis synovial fibroblasts  Download fulltext PDF



Autor: Thomas Zimmermann - Elke Kunisch - Robert Pfeiffer - Astrid Hirth - Hans-Detlev Stahl - Ulrich Sack - Anke Laube - Eckehard

Fuente: https://link.springer.com/







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