GSM8636265: IL-15SA; Mus musculus; RNA-Seq
Tumors were dissected and dissected tumors were mechanically minced and digested through stirring in RPMI 1640 medium that contained collagenases Type IV (1 mg/mL, Thermo Fisher Scientific), dispase II (100 μg/mL, Sigma-Aldrich), hyaluronidase (100 μg/mL, Sigma-Aldrich), and DNase I (100 μg/mL, Sigma-Aldrich) at 37 °C for 60 min. Red blood cells were removed with ACK lysing buffer after digestion. TILs were enriched by density gradient centrifugation against Percoll (GE Healthcare) and resuspended in FACS buffer (1 × PBS containing 0.2 % BSA (wt / v), Sigma-Aldrich), then sorted by MACS (Miltenyi) and FACS (Sony). CD45.2+CD3-NKp46+ NK cells were sorted as described above, cells from the same treatment group were pooled together and resuspended at 3 × 105 cells/mL in PBS. Next, 100 µL cell resuspension was loaded onto a standard GEXSCOPE® microwell chip. scRNA-seq library was constructed using the GEXSCOPE® Single-Cell RNA Library Kit according to the manufacturer's instructions (Singleron Biotechnologies GmbH). Individual libraries were diluted to equimolar and pooled for sequencing. Pools were sequenced on a lane of an S4 flow cell on an Illumina NovaSeq 6000 with 150 bp paired-end reads to a total depth of 90 GB per sample and an average read depth per cell of 50,000 reads. Raw reads were processed with FastQC to evaluate the quality of raw data.