GSM9039359: B16F10_tumor, CD45+, Pmel-1_CTL_transfer, day_3; Mus musculus; RNA-Seq
For preparation of tumor-infiltrating immune cells, tumor tissues were cut into small pieces and incubated in RPMI-1640 medium containing 0.2% collagenase (Fujifilm Wako Pure Chemical Corporation) and 2 KU/mL DNase I (Sigma-Aldrich) at 37 °C for 40 minutes. The digested tissues were passed through a 70 μm cell strainer. Single-cell suspensions were stained with anti-mouse CD45 antibody and viable CD45⁺ cells were isolated using a cell sorter. Equal numbers of CD45⁺ cells from three mice were pooled for each time point and used for single-cell RNA sequencing. Single-cell suspensions were loaded onto the 10x Genomics Chromium Controller using the Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (Dual Index), following the manufacturer's instructions. After GEM generation and reverse transcription, cDNA was amplified and sequencing libraries were constructed. Libraries were quantified using Kapa library quantification kit (Roche).