GSM5860222: Combination anti-PD-1 and anti-PSGL-1 treated tumor-infiltrating immune cells; Mus musculus; RNA-Seq
Tumors were dissected at 18dpi and processed using tumor digest enzymes A and D in RPMI on the GentleMacs tumor processcor setting 37C_h_TDK_1. Tumor were then passed through 70 micron strainer with sterile wash media (HBSS with 1%FBS, 0.5% HEPES, and 2%PSG). Tumors were spun and resuspended in sterile PBS with 0.04% BSA. Tumor cells were counted and stained with PI for 10 min on ice at 25x10^6 cells/mL. Cells were washed with sterile PBS +0.04% BSA and then stained for CD45.2 (1:300 dilution) with FC block (1:100) at 25x10^6 cells/mL for 20 min on ice. Cells were washed 3x and resuspened in sterile PBS + 0.04% BSA and brought to the flow core at 10x10^6 cells/mL for flow sorting. Purified PI- CD45.2+ cells were sorted for each treatment group. Sorted cells were spun and resuspenede at a concentration of 700 cells/uL. RNA libraries were prepared using the 10X Genomics Chromium Single-Cell Platform protocol scRNA-seq