Fig. 1: High-throughput screening platform to evaluate bsAb-induced T-cell-mediated tumoroid killing. | Communications Biology

Fig. 1: High-throughput screening platform to evaluate bsAb-induced T-cell-mediated tumoroid killing.

From: CD3-engaging bispecific antibodies trigger a paracrine regulated wave of T-cell recruitment for effective tumor killing

Fig. 1

A Workflow for bsAb screening: tumoroids are printed at identical x-y-z positions in 96-well plates prefilled with collagen gel, followed by the addition of T-cells and bsAbs on top of the gel. Migration and recruitment of T-cells (CMFDA; green) to tumoroids (Hoechst; blue) and tumor killing (PI; red) are monitored using automated confocal microscopy at different time points. The cartoon of the 96-well plate was obtained from Servier Medical Art (https://smart.servier.com/), which is licensed under a Creative Commons Attribution 4.0 Unported License (https://creativecommons.org/licenses/by/4.0/). B Schematic representation of the automated image analysis procedure: stacks of confocal images are captured at 10 μm vertical spacing throughout the z-axis of the tumoroid. Tumor nuclei and T-cells are identified separately and tumor nuclei with or without overlapping PI signal are counted in each z-section. Tumor killing is determined by calculating the percentage of PI positive tumor cells in the entire tumoroid. T-cell recruitment is determined as the sum of all T-cells within the tumoroid counted in each z-section. PI Propidium iodide.

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