GSM7844334: G3 scRNA-seq; Mus musculus; RNA-Seq
MC38-HER2 bearing mice were sublethally lymphodepleted by total body irradiation (4 Gy) on day 5, followed by i.v. adoptive transfer of HER2 CAR-T or IL-10 HER2 CAR-T cells on day 6. On day 18, mice were sacrificed, and CAR-T cells were isolated from tumors or spleens as described above. NSG mice were inoculated (s.c.) with PANC1-CD19 cells (2 × 106) and received i.v. adoptive cell transfer of CD19 CAR-T or IL-10 CD19 hCAR-T cells (1 × 106) on day 8. On day 39, mice were sacrificed, and CAR-T cells were isolated from spleens as described above. Sorted HER2 CAR-T cells from tumors were then pelleted for 5 min at 400 relative centrifugal force and washed once in PBS with BSA (0.04% wt/v). Afterward, HER2 CAR-T or IL-10 HER2 CAR-T cells were suspended in 100 μl of Cell Multiplexing Oligo (LMO, 10x Genomics), respectively. LMO-barcode labeling was performed for 10 min at room temperature, and then cells were washed twice with PBS, resuspended in PBS with BSA (0.04% wt/v), filtered, and pooled. Labeled cells were subjected to single cell encapsulation using a Chromium Single Cell Instrument and reagents. Sorted CD19 hCAR-T cells or IL-10 CD19 hCAR-T cells from spleens were subjected to single cell encapsulation using a Chromium Single Cell Instrument and reagents separately. For G1and G2, A Chromium Next GEM Chip G was loaded with the appropriate number of cells and the sequencing libraries prepared using 10x Genomics reagents according to the manufacturer's instructions and passed quality control. For G3, cells were subjected to single cell partitioning using microchip, mRNA Capture and sequencing libraries were prepared using GEXSCOPE Single Cell RNA Library Kit Cell V2 reagents (Singleron Biotechnologies GmbH) according to the manufacturer's instructions and passed quality control.