GSM7596310: primary FNH, scRNA-seq; Homo sapiens; RNA-Seq
Freshly excised tissue was washed with RPMI-1640 medium (Thermo Fisher Scientific), placed on ice, and cut into 1-2 mm pieces. The samples were then incubated at 37˚C for 40 minutes in a digestion solution composed of 0.25% trypsin (Thermo Fisher Scientific.) and 10 µg/ml DNase I (MilliporeSigma) dissolved in 5% fetal bovine serum (FBS; Thermo Fisher Scientific). Manual oscillation of the samples was performed every 5 minutes, and the resulting cell suspension was filtered twice using a 40-µm nylon mesh (Thermo Fisher Scientific). Erythrocytes were removed using 1X Red Blood Cell Lysis Solution (Thermo Fisher Scientific). Subsequently, the samples were resuspended in phosphate-buffered saline (PBS) containing 0.08% bovine serum albumin (BSA; MilliporeSigma) and filtered through a 40µm cell filter (MilliporeSigma). A single-cell 3'-library was constructed on the Chromium platform, following the manufacturer's instructions. The samples were processed utilizing the 10× Genomics V2 barcoding chemistry kit.