GSM8647666: HCC_T, scRNA-seq; Mus musculus; RNA-Seq
Tissues were finely minced and subjected to enzymatic digestion at 37°C for 30 minutes with gentle shaking at 200 rpm in a digestion buffer. After digestion, cell suspensions were filtered through a 100 μm cell strainer and subsequently washed with 1× phosphate-buffered saline (PBS) containing 2% FBS and 2 mM EDTA. The washed cell suspensions from liver/tumor were further applied for immunocyte purification by using 35% Percoll. Red blood cells were removed by Red Cell lysis buffer. Afterwards, the cells were then stained with fluorochrome-conjugated surface antibodies in 1× PBS containing 2 mM EDTA. Viable cells were identified by using Live/Dead Fixable Viability Dye. Cell sorting from mouse liver/tumor tissues was performed on BD FACSAria, and the viable CD45+ cells (immunocytes) were sorted for scRNA-seq. Libraries were prepared with Chromium Next GEM Single cell 3' Reagent v3.1 Kits according to the manufacturer's protocol.