GSM8769381: TILs from NSG CD8ab CD4 and CD8 combo; Homo sapiens; RNA-Seq
Dissected tumor from NSG models were digested with tumor dissociation kit(Miltenyi), washed and stained in FACS buffer, then sorted with BD Symphony S6 flow cytometry sorter. CD4+ and CD8+ T cells obtained were then proceeded with 10X library prep. Cryopreserved tumor single-cell suspension from in vivo murine experiment were thawed, washed and processed using a 10x Chromium Controller following the 5' Chromium Single Cell V(D)J Reagent Kit manual (10x Genomics). Library preparation was carried out according to the manufacturer's protocol without modifications. Library quality was assessed using TapeStation High Sensitivity (Agilent) to evaluate library size, Qubit (Thermo Fisher) to measure dsDNA quantity, and KAPA qPCR analysis (KAPA Biosystems) to determine the quantity of amplifiable transcript. Samples were pooled in equimolar proportions and sequenced on an Illumina HiSeq 2500 in rapid run mode, following the standard 10x Genomics protocol. TCR target enrichment, 5' gene expression library preparation, and TCR library construction were performed using to the 5' Chromium Single Cell V(D)J Reagent Kit manual (10x Genomics).