GSM8848849: Mouse KPC tumor, intratumoral VIVIT, rep2; Mus musculus; RNA-Seq
Tissues were minced and digested using a 1:1 ratio of Collagenase Type IV (Cat#17104019; Thermo Fisher) and Collagenase Type II (Cat#17101015; Thermo Fisher) at a concentration of 1500 U/mL in DMEM (Cat#10569010; Thermo Fisher) for 90 minutes at 37°C. Tissue pieces were agitated at 150 rpm during enzymatic digestion using an orbital shaker. Collagenase activity was stopped using DMEM with FBS, and digested tissue was strained through 100 μm (Cat#4620E99; Corning) and 40 μm cell strainers (Cat#4620F05; Corning). Cells were centrifuged at 1500 rpm / 524 rcf for 5 minutes at 4°C resulting in a single-cell suspension. Samples were resuspended with 0.04% UltraPure BSA (Cat# AM2618; Thermo Fisher). scRNA-seq libraries were made following the 10x Genomics Single Cell 3' Reagent Kits User Guide (v3 Chemistry) for control1 and vivit1. The remaining 4 libraries were made following the 10x Genomics GEM-X Universal 3' Gene Expression v4 4-plex Reagent Kits User Guide.