Next Generation Sequencing of Chimeric antigen receptor T (CAR-T) cells polarized under Th9 condition

Isolated human T cells was stimulated by anti CD3/CD28 Dynabeads and polarized under Th1 (IL2) or Th9 (IL4 plus TGF beta) condition.After 24 hours, cells were transduced with lentivirus. Medium were renewed every 2 days and cells were maintained at a concentration of 1 million per milliliter. On day 16 of culture total RNA was isolated from CAR-T cells using RNeasy Mini kits (Qiagen) following the manufacturer's instruction. Quality of total RNA was evaluated using RNA 6000 Nano LabChip. Qualified RNA was subject to next generation sequencing. Next ggeneration sequencing was performed on chimeric antigen receptor T cells polarized under Th1 or Th9 condition.