GSM7901274: Glioma_8_PBMC_TIL_CITEseq; Homo sapiens; OTHER
Peripheral blood mononuclear cells (PBMC) were isolated from whole blood by Ficoll-Hyapque density gradient centrifugation. Freshly resected glioma tumors were mechanically disrupted and the resulting tissue fragments were digested with 50 mg/mL of Liberase DL  in 5 mL of serum-free RPMI for 15 minutes at 37°C. Live CD45+ (i.e. all live immune cells) were sorted from PBMC and glioma tissues. Single-cell RNAseq was performed using 10X Genomics and libraries were sequenced on Illuman NovaSeq6000 instruments. The library generation strategy was as per the manufacturer's instructions for either 10X Genomics 3' or 5' based chemistries for a given sample. For some samples, cell hashing was performed to run the PBMC and tumor infiltrating leukocytes (TIL) in the same lane of the 10X Chip. For these samples, oligonucleotide-labeled antibodies were used to label PBMC and TIL and as specified for each sample. Cell hashing libraries were generated as per the 10X Genomics manufacturer's instructions.