GSM7969080: CD45+ TILs from mouse 3 treated with aPD-L1+E.colipApyr; Mus musculus; RNA-Seq
Tumors were cut in small pieces and resuspended in RPMI1640 with 1.5 mg/mL Type I Collagenase (Sigma), 100 μg/mL DNase I (Roche), and 5% FBS, digested for 45 minutes at 37°C under gentle agitation. The digestion product was then passed through a 70 μm cell strainer to obtain a single-cell suspension. Cells were stained with Zombie Live/Dead and CD45 and immune cells were sorted using BDFACSAria and immediately processed for scRNA-Seq scRNA-seq was performed immediately after FACS sorting of CD45+ TILs with the Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1(10X Genomics, Pleasanton, CA) following the manufacturer's protocol. Target number of captured cells ranged from 2,000 to 10,000 cells. Sequencing libraries were prepared per manufacturer's protocol.