GSM8995790: LNP-P20-mAb_TCR_1; Mus musculus; OTHER
Tumors were harvested and enzymatically dissociated into single-cell suspension using the Mouse Tumor Dissociation Kit (Miltenyi Biotech, 130-096-730) in combination with the gentleMACSTM Octo Dissociator with Heaters (Miltenyi Biotech). Following dissociation, red blood cells were lysed, and cell suspensions were filtered through a 40 μm cell strainer (Corning) and resuspended in RPMI-1640 medium supplemented with 5% FBS. Viable leukocytes were enriched by staining with APC-Cy7-conjugated anti-mouse CD45 antibody following Fc receptor blockade, and dead cells were excluded using Zombie Aqua Fixable Viability Kit. CD45+ immune cells were sorted using a FACSAria II flow cytometer (BD Bioscience). For each sample, approximately 10,000 sorted CD45+ cells were processed for library construction using the Chromium Next GEM Single Cell 5' Reagent Kits v2 (10× Genomics, PN-1000263). In parallel, T cell receptor (TCR) libraries were prepared using the Chromium Single Cell V(D)J Enrichment Kit.