IncuCyte™ Poster Snapshots - Kinetic Live Cell Imaging Assays for Immune Cell Killing



July 27, 2016

Presented by Nadezda Rodionova at ITOC3, March 2016. ITOC3 is the third edition of the Immunotherapy of Cancer Conference held at the Munich Tumour Center, Munich, Germany, from 21 to 23 March 2016 N. Rodinova, N. Bevan, C. Szybut, T. O’Callaghan, T. Dale & D. Trezise. Essen BioScience, Welwyn Garden City, Hertfordshire AL7 3AX, UK View the full poster: ------------------------------------ Summary and Impact: • Cancer immunotherapy offers the potential for specific eradication of tumour cells and the prevention of cancer recurrence. • Here we describe fully kinetic live cell imaging assays to quantify immune cell mediated killing of adherent and suspension tumour cells in co-culture. • RFP-nuclear labelled tumour cells were seeded either overnight in 96 well plates or at time of assay in ultralow attachment U-bottom plates. Immune cells (PBMCs, isolated CD8+ T or NK cells) were then added to the wells in combination with various activators and a non-perturbing caspase 3/7 apoptosis green fluorescence detection reagent. • Phase and dual-colour fluorescence images were captured every 2h for 4-7 days using IncuCyte® ZOOM. • Tumour cell numbers were quantified by direct nuclear object counting (RFP). The number of apoptotic tumour cells were measured using green (caspase 3/7) object counting and size/brightness filtering to remove dying immune cells. • This method is illustrated and validated with anti-CD3 and IL-2 mediated cell killing and antibody driven cell killing both with adherent and suspension target cells. • These validation data demonstrate that live cell imaging can be used to discern the full time course and specificity of immune cell killing, without the need to lift cells, use Ab labels or radioisotopes (e.g. Cr51). • HD images and time-lapse movies facilitate a clear understanding & compelling verification of the underlying biology.

Cell ScienceDrug DiscoveryImaging/Microscopy

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