GSM5203286: CONTROL_DC; Homo sapiens; RNA-Seq
Liver biopsies from bariatric surgery patients were collected in RPMI (Supplemented with 10%FBS) and immediately transported on ice to the laboratory. A section of tissue was formalin fixed for histopathological evaluation; however, the bulk of the tissue was digested using Miltenyi Tumor dissociation Kit (cat no. 130-095-929) as per the manufacturer's instructions. Single cell suspensions thus generated were stored in Liquid nitrogen in FBS+20% DMSO until further processing. For sorting and scRNAseq of DCs obtained from control and NASH livers, the stored cells were thawed in a 37oC water bath, washed once with complete media and once with FACS buffer (PBS+2% FBS). After FC blocking, cells were stained for 30 min and washed once with FACS buffer and once with sorting buffer (PBS+0.5mM EDTA). Sorting was carried out at DKFZ cytometry facility using BD FACS Aria. Bulk-sorted CD45+ Lin- CD11c+ were then processed for 10X (10X Genomics). 10x v.3; Single cell RNA-seq 3' : 28bp R1 and 91 bp R2