GSM7851056: LN3, scRNAseq; Homo sapiens; RNA-Seq
Fresh lymph node biopsies from patients with newly diagnosed (13) or relapsed/refractory (1) cHL, as well as reactive lymphoid tissues (5) from patients were utilized in this study. To prepare cHL tissues for downstream analyses, they were cut into small pieces and incubated in 1 mg/ml collagenase IV (Sigma-Aldrich) and 20 mg/ml DNAse I (Roche) at 37 °C for 30 min and then passed through a 100 micron filter to generate single cell suspensions, which were frozen for future analysis. B-cell depletion of each sample was achieved using CD19-biotin and anti-biotin microbeads (Miltenyi) and negative selection to enrich T cell populations. A total of 8,000-10,000  cells per sample were processed using a Chromium Next GEM Single Cell 5' Kit v1.1 to generate cDNA libraries, and sequencing was performed using the 10x Genomics Chromium Single Cell 5' Gene Expression workflow for Single Cell RNA Sequencing. Single-cell libraries from four samples were pooled and sequenced on one NovaSEQ SP 100 base PET lane to achieve an average sequencing depth of 2,981 reads per cell.