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

Citations Over TimeTop 10% of 2000 papers

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.

Related Papers

• → Navigating challenges: optimising methods for primary cell culture isolation(2024)35 cited
• → Primary Cell Cultures of Drosophila Cells(1997)1 cited
• Analysis different methods between amniotic fluid cells primary culture and passage culture in prenatal diagnosis(2013)
• → Assay Development Using Primary and Primary- Like Cells(2009)
• → P22 Classification and characterisation of primary batteries(1997)