I used to perform the cytotoxicity assay. The short protocol is like that first I labeled the target cell with CFSE, Co-culture with CAR-t and then next day stained with PI and analyzed by FACS.
How can we calculate the %cytotoxicity? According to online protocol, % cytotoxicity= Dead target cell/Dead*Live cells. somewhere in literature, Percent cytotoxicity of target cells is measured by comparing survival of target cells relative to the survival of negative control cells. At multiple places, I found different methods for the calculation of % cytotoxicity.
Kindly please recommend me the most widely usable and acceptable method for calculation of % cytotoxicity??
Thanks.
Dear Waqas Navas
in my opinion, the most cemprehensive approach would be to express %cytotoxicity as dead target cells / (dead+live target cells). This will additionally offer you the possibility to directly compare %cytotoxicity between different effector cell/target cell ratios.
best wishes
Andreas
@ Andreas Benesic Okay i got it. But i have 2 questions here based on the above mentioned formula. 1. The dead cells which you mention are the total dead cells (Effector+ targets) ? 2. and Whats about the (-CFSE and -PE) unstained cells, should we ignore that quadrant in our calculation?
Thanks.
Dear Waqas Navaz
I would recommend doing the calculation for Target cells (CFSE pos) and for effector cells (CFSE neg) separately
Best wishes
Andreas
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