title: 1607_SF
organism: Homo sapiens
description: Protocols: Blood and synovial fluid samples were collected with written informed patient consent from PsA patients undergoing intra-articular aspiration at Oxford University Hospitals. The study was performed in accordance with protocols approved by the Oxford Research Ethics committee (Ethics reference number 06/Q1606/139). Mononuclear cells were immediately isolated by density centrifugation then stained by a panel of fluorescently conjugated antibodies in staining buffer (RNAse-free PBS, 2mM EDTA). The following antibodies were used: CD3-FITC (SK7), CD4-APC (RPA-T4), CD8a-PE (RPA-T8), CD45RA-BV421 (HI100) (all BioLegend), together with eFluor780 viability dye (eBioscience). Stained cells were sorted into memory-enriched (CD45RA negative) CD3+CD4+CD8- or CD3+CD8+CD4- fractions, which were then combined in a 1:1 ratio before loading onto the 10x Chromium Controller. A separate channel was used for the synovial and blood derived cells from each patient. Standard 10x protocols for the kit were used to load cells into droplets containing barcodes. Cells were captured and cDNA preparation performed according to the Single Cell 5′ Protocol recommended by 10x Genomics.