GSM9011720: Lung adenocarcinoma, tumor tissues, patients#2; Homo sapiens; RNA-Seq
Tissues were rinsed with ice-cold PBS three times to remove blood clots and then minced on ice. The tissue block was incubated with a digestion solution made of 0.25% trypsin and 10 µg/mL DNase I, at 37°C with a shaking speed of 50 rpm or 30 min. Cell suspensions were filtered using a 70-μm cell strainer and then through a 40-μm strainer again, and the filtrate was collected into a new centrifuge tube. The cell pellet was resuspended in 3 ml of red blood cell lysis solution and incubated on ice for 5 min. Then, 10 ml of precooled PBS was added and centrifuged at 250 × g for 5 min. The cell pellet was resuspended in PBS containing 0.5% FBS. Cells were stained with 0.4% AO/PI to evaluate cell viability. 10 × Genomics single-cell capture, Illumina library construction were performed by Capital Bio-Tech (Beijing, China).