title: Single cell RNA sequencing of paired PBMC and synovial tissue cells from Rheumatoid Arthritis (RA) patients naive to treatment
design: Single cell RNA sequencing of paired PBMC and synovial tissue cells from Rheumatoid Arthritis (RA) patients naive to treatment
library_construction_protocol: To investigate synovial tissue myeloid DC and T-cell heterogeneity in RA, we performed scRNAseq on matched peripheral blood mononuclear cells (PBMC) and synovial tissue (ST) biopsies (n=3). Synovial biopsies were taken using 14G Precisa Needle (HS Hospital Service, Italy) in ultrasound-guided protocol, and digested as described previously. All live cells or CD45pos immune cells, were FACS sorted, after exclusion of dead cells with Fixable Viability Dye eFluor® 780 (eBiosciences). Maximum 20,000 immune cells from blood or tissue cells were sorted into Protein LoBind 1.5 ml Eppendorf tube containing 300 μl of RPMI media with 10% of FSC. Cells were loaded onto a Chromium Controller (10X Genomics) for single-cell partitioning, followed by library preparation using Single-Cell 3' Reagent Kits v2.  As recommended by 10X Genomics protocol for the 10X Single Cell 3' Reagent Kit v2 As recommended by 10X Genomics protocol for the 10X Single Cell 3' Reagent Kit v2