GSM9009159: TCR_isotype_2; Mus musculus; OTHER
TIL isolation, tumours were harvested on day 21 or 22, cut into small pieces and digested with the Tumour Dissociation Kit (Miltenyi Biotec 130-096–730) for 40 min at 37 °C in accordance with the manufacturer's protocol. Cell suspensions were obtained by crushing and passage through 100 μm-mesh cell strainers. Red blood cells were removed with 1x Red Blood Cell Lysis Solution  (Miltenyi Biotec 130-094-183). The cells were washed twice with 2% FCS in PBS and for the preparation was enriched in CD8 and CD4 cells with the MultiMACS system (Miltenyi Biotec). Cells were labeled with anti-CD8a or anti-CD4 (L3T4) antibody-conjugated microbeads (Miltenyi Biotec 130–117–044 or 130-117-043, respectively) and then purified with the POSSEL program on MultiMACS. CD3+ TIL were loaded onto a Chromium Single-Cell Chip (10x Genomics), according to the manufacturer's instructions, for co-encapsulation with barcoded Gel Beads at a target capture rate of ~10,000 individual cells per sample. Captured mRNAs were barcoded during cDNA synthesis with Chromium Next GEM Single-Cell 5' GEM, Library Kit v2 (10X Genomics) according to the manufacturer's instructions.