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Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery
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Capitoli
- 00:12Introduction
- 01:07Setting up the JIMT-1 Enhanced Fluorescent Protein (EGFP) Spheroid Model
- 03:43Coating the HCS Plate
- 04:33Transfer of Spheroids to the HCS Plate
- 04:46Pre-treatment of JIMT-1-EGFP Spheroids with Test Compounds
- 05:14Induction of Antibody-Dependent Cellular Cytotoxicity (ADCC) by Adding the Effector Cells
- 07:10Annexin V-647 Staining of Spheroids
- 07:34Imaging
- 11:43Representative Results
- 12:52Conclusion
Here, we present a method to identify compounds that modulate the ADCC mechanism, an important cancer cell-killing mechanism of antitumor antibodies. The cytotoxic effect of NK cells is measured in breast cancer cell spheroids in the presence of Trastuzumab. Image analysis identifies live and dead killer and target cells in spheroids.
