GSM8018832: Vagatomized Liver 2; Mus musculus; RNA-Seq
Eight samples were prepared for scRNA-seq. Solid RIL175 tumors were excised from SV and HV livers (n = 3 / group) 21 d following tumor initiation.  RIL175 tumor-bearing mice (n = 3 SV, 3 HV mice) from non-tumorous liver and matched (pooled) tumors. Immune cells from tumors and livers were isolated as described in earlier methodology. Following surface staining, Live+CD45+ cells were sorted into PBS + 0.04% BSA (ThermoFisher Scientific, catalogue #B14) at the NIH Flow Cytometry Core using the Aurora CS Cell Sorter (Cytek® Biosciences, RRID:SCR_XXXXX).  Subsequent cell counts and viability were determined with propidium iodide/acridine staining using the Luna FL Automated Fluorescence Cell Counter (Logos Biosystem). Samples were provided to SCAF (NIH Single Cell Analysis Facility) for library preparation,  sequencing, and data processing. Cell counts were determined via LunaFx7 fluorescent cell counter (Logos Biosystem) and adjusted cell counts targeting 6,000 cells/lane were prepared. Library preparation and sample loading were performed by SCAF and in accordance with the 10X Genomics Single Cell User Guide utilizing Chromium Connect (10X Genomics automation platform).    Sequencing was performed using the NovaSeq 6000 Reagent Kit (Illumina). Two sequencing runs (NovaSeq 6k S1 100-cycle) were performed via 5' Immune Profiling v2 Dual-Index chemistry (10x Genomics).