GSM8825426: T cells from Synovium tissue, TCR, AxSpA patient, rep 1; Homo sapiens; RNA-Seq
Synovial tissue was obtained from surgical procedures. Cells were isolated from fresh tissue through enzyme digestion using 100 µg/ml Liberase TL and 100 µg/ml DNase in RPMI. The isolated cells were either directly subjected to the Chromium Single Cell Gene Expression Chip (10X Genomics) assay or sorted into live CD3+ and CD3- populations using Fluorescence-activated Cell Sorting (FACS). For FACS, the single cell suspension was first incubated with Human TruStain FcX solution (Biolegend) to block Fc receptors and then stained with anti-CD3 (clone OKT3) and Fixable Viability Dye eFluor 780 (eBioscience/ThermoFisher). Approximately 20,000 cells from each population were loaded onto a Chromium Next GEM Chip (10X Genomics), and scRNA-seq libraries were prepared according to the manufacturer's instructions. For samples directly applied to library preparation without sorting, around 20,000 cells from each sample were loaded. The resulting DNA libraries were sequenced at >50,000 reads per cell on a NovaSeq 6000 using PE150 mode.