A Real-Time In Vitro Cytolysis Assay to Evaluate the Potency of CAR T Cells
Transcrição
Begin with a cell culture medium-containing E-Plate — a plate with gold microelectrodes in each well's bottom.
Seed the plate with target cancer cells, adhering to the gold microelectrodes, forming a physical barrier between the electrodes.
Transfer the plate to a cell analyzer and apply an electrical signal.
Adhered cells disrupt the electron flow between the electrodes, resulting in an impeded flow of electrons, measured as impedance.
As these cells proliferate and adhere to the electrodes, electrical impedance increases proportionally with the adherent cell number, enabling real-time measurement.
Introduce chimeric antigen receptor T cells or CAR T cells expressing chimeric antigen receptors. These receptors interact with cancer cells' antigens, activating T cells, and forming an immunological synapse.
Activated CAR T cells release cytotoxic substances, including perforins and granzymes.
Perforins form pores in the cancer cell membrane, facilitating granzymes to enter the cells and induce cytolysis.
This promotes cell detachment, decreasing electrical impedance, correlating with CAR T cells' cytolysis potential.
