A CARTcelltherapy for advanced ovariancancer developed cityofhope NatureComms
, or phenol red containing 2% FBS and 1 × AA). Cells were incubated with primary antibodies for 30 min at 4 °C in the dark. For secondary staining, cells were washed twice prior to 30 min incubation at 4 °C in the dark with either Brilliant Violet 510 , Brilliant Violet 570 , Brilliant Violet 605 , Brilliant Violet 650 , fluorescein isothiocyanate , phycoerythrin , peridinin chlorophyll protein complex , PerCP-Cy5.
for the full antibody list. Data was acquired on a MACSQuant Analyzer 16 cytometer and analyzed with FlowJo v10.8.For tumor cell killing assays, CAR T cells and tumor targets were co-cultured at indicated effector:tumor ratios in complete X-VIVO or cRPMI without cytokines in 96-well plates for the indicated timepoints and analyzed by flow cytometry as described above. Tumor cells were plated overnight prior to the addition of T cells.
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