GSM8421835: CAR-T_pre_WT; Homo sapiens; RNA-Seq
On day 12, the CD8α+ and CAR+ cells sorted by a cell sorter (Sony) were used as an RNA sequencing of pre-sample. Regarding post-samples, T cells were subjected to the repeated stimulation assay in 6-well plates, transferring T cells to freshly seeded cancer cells (A549) every 48-72 hours. Cultured T cells were sorted after chronic stimulation using cell sorter (Sony) to obtain a pure population of HER2 CAR positive CD8α+ T cells from the co-cultures with target cancer cells. Total RNA from pre and post-CAR T cells was harvested using RNeasy mini kit (Qiagen). RNA libraries for RNA-seq were prepared using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB E7490) and NEBNext Ultra RNA Library Prep Kit for Illumina (E7530) following manufacture's protocol