GSM7968900: Ctrl replicate3 scRNA-seq; Mus musculus; RNA-Seq
Tumor tissue was incubated in dissociation buffer containing 2 mg/ml Collagenase Ⅳ(sigma,C5138), 2 mg/ml Collagenase II sigma C6885， 10 mg/ml Dispase II (corning,354235) and 50,000U/ml DNase I (sigma, DN25) for 30 min at 37 °C. Enzymatic digestion was quenched with Dulbecco's Modified Eagle Medium (DMEM, Gibco) supplemented with 10% fetal calf serum (FCS, 10270106, Gibco). Cells were resuspended at a concentration of 1 million cells per ml and stained with PBST containing 1% FCS. Then 15 µl of single-cell suspension at a concentration of ~900,000 cells/ml was loaded into one channel of the ChromiumTM Single Cell B Chip (10X Genomics，1000073), aiming for a recovery of 8000–9000 cells. The Chromium Single Cell 3′ Library & Gel Bead Kit v3 (10X Genomics, 1000075) was used for single-cell barcoding, cDNA synthesis and library preparation, following the manufacturer's instructions according to the Single Cell 3′ Reagent Kits User Guide Version 3.