Transposon system-based manufacture of CAR-T cells via electroporation
By using transposon systems like the Sleeping Beauty (SB)5,6 system or piggyBac7 combined with culturing on artificial Antigen Presenting Cells (aAPCs), it is possible to achieve stable integration and the selective proliferation of CAR-T cells.
As a brief overview of the procedure, T cells are collected and transfected using electroporation, with a DNA plasmid carrying an IR/DR flanked CAR transposon as well as a DNA plasmid encoding a transposase. The mixed population of untransfected T cells as well as successfully transfected CAR-T cells with the CAR expression cassette integrated is then cultured on APCs (e.g. K562 cells that express endogenous molecules for T cell activation and proliferation). Thus, only the CAR expressing T cells are selected for re-infusion into the patient’s body.
Alternatively, electroporation of CAR mRNA has been investigated. This method enables a high level of CAR expression for a limited time of 3-5 days. This ensures an important safety feature for patients by “testing” for severe side effects before treating long-term.
Regardless of whether viral transduction or electroporation is used, adoptive immunotherapy via CAR-T cells is a huge focus area and may lead to a powerful tool to fight not only cancer, but also infectious diseases. There is still a lot of research needed to see CAR-T cell immunotherapy to its full potential. Lonza is committed to supporting your CAR-T cell research. That’s why we offer the
NucleofectorTM Technology and corresponding T cell Nucleofector
TM Kits for efficient T cells transfection. In addition, we provide human PBMCs as well as purified CD4+ T cells from a variety of different donors. Please contact
Scientific Support for more information.