Evaluating CAR-T Cell Cytotoxicity Using LDH Assay

LDH based assay was conducted to evaluate the cytotoxic effects of CAR-T cells [30 (link)]. Tumor cells (1 x 10⁵ cells /well) were added to the 96-well plates, and then according to the effect of target ratio 1:1, 5:1, and 20:1, c-Met CAR-T, CD19 CAR-T, and non-transduced T cells were added. Next, the incubation was performed at 37°C and CO₂ (5%) post 6 hours of culture.And LDH detection reagent was added (C0017, Beyotime, China), the optical density was recorded at 490 nm wavelength.
The mean value of the three wells was substituted into the following formula to calculate the killing efficiency % = (the measured hole – effector cells natural release OD value – target cells natural release OD value + blank control OD value)/(the maximum release hole OD value of the target cells – target cells natural release OD value).