GSM9535805: P063, Liver metastasis, PBMC and healthy liver, Tconv, ADT; Homo sapiens; OTHER
PBMCs were isolated from heparinised blood by density centrifugation using Ficoll-Paque PLUS (Sigma-Aldrich). Tissue samples obtained from different surgical liver resections (HCC and paired non-tumoral background tissues from HCC resections; and histologically healthy liver tissue from liver metastasis resections) were collected in cold DMEM medium (Life Technologies) and processed within 2 hours. Tumour and tissue single cell suspensions were obtained by enzymatic digestion with the BD HorizonTM Dri Tumour & Tissue Dissociation Reagent (TTDR) (BD Biosciences) as per manufacturer instructions. scRNAseq libraries (5´ gene expression library GEX, T cell receptor enriched library VDJ, and surface protein library containing the TotalSeq-C CITE-seq antibodies) were prepared using the Chromium Single Cell Immune Profiling kit (10x Genomics, v1.1) according to the manufacturer's instructions. The generated scRNAseq libraries were sequenced using a total of 100 cycles (paired end reads) with a NovaSeq 6000 S2 flow cell (Illumina).