GSM7034607
Anti-mesothelin M5 CAR-TCONV and CAR-TRM cells were harvested after nine days of in vitro expansion. Dead cells were eliminated using the Dead Cell Removal Kit (Miltenyi Biotec). Mesothelin-directed M5 CAR-TCONV and -TRM cells were magnetically isolated using anti-mesothelin-coated beads. After isolation, T-cell viability and quantity were evaluated using trypan blue exclusion and cells were resuspended in PBS containing 0.04% BSA. Approximately 20,000 cells were loaded per reaction to capture ~10,000 cells for scRNA-seq library preparation using Chromium Single Cell 5' v2 Reagent kits (10X Genomics) according to manufacturer's protocol. Sequencing was performed on a NovaSeq 6000 instrument.