Presenting Author: Shannon McGettiganAdditional Authors: Yanping Luo, Keisuke Watanabe,
John Scholler, Carl H June Research Specialist University of Pennsylvania Cytotoxicity
assays are an important characterization in the development of anti-cancer therapeutics. Chromium release assays are considered the gold standard for
evaluating lymphocyte cytotoxic activity but requires the burden of using radioactive
materials, is time consuming, and is limited to a single time point.our lab and others
have developed flow cytometric and luciferase based cytotoxcity assays for screening
and evaluating novel therapeutic CAR T cells against a wide range of cancer cell lines
and primary tumor; however, these can also be time consuming and limited to a single
snapshot. In order to monitor the overall killing activity of our CAR T cell therapies as
a function of time and more rapidly drive our understandings in early development of
therapeutic T cells, we have started to utilize the xCELLigence real time cell
analyzer (RTCA). Our studies have compared how measurement of changes in
adherent cell’s electrical impedance compares to our standard cytotoxicity
measurements by the remaining viable cell numbers in flow based assays or relative
changes in luciferase activity. We found that a correlation exists between the platform
for measuring cytotoxicity. Real time cellular impedance analysis reveals kinetic
differences that cannot be captured practically with conventional fixed end point
platforms. We have found that using the xCELLigence platform has many benefits
beyond just real time monitoring. The assay requires minimal number of cells which
can be retrieved for further analysis, saves time, provides a kinetic readout of
combination therapies, and is a quick quality control cytotoxicity assay for
therapeutic T cells used in in vivo experiments. Together the xCelligence Real time
platform shows valuable utility for screening gene modified T cells cytotoxic function,
characterizing the kinetics of their activity, evaluating the dosage and timing of
combination therapies in vitro and providing a quick stable platform for quality control
of therapeutic T cells.
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