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Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery
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JoVE Journal Cancer Research
Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery

Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery

DOI:
10.3791/65922-v

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13:19 min

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April 26, 2024

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, , , , ,
1Department of Medical Chemistry, Faculty of Medicine,University of Debrecen, 2Doctoral School of Molecular Medicine,University of Debrecen, 3National Academy of Scientist Education, 4HUN-REN-DE Cell Biology and Signaling Research Group

Chapters

  • 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

Summary

Automatic Translation

Automatic Translation

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.

Tags

Antibody-dependent Cellular CytotoxicityADCCTumor Spheroid ModelDrug DiscoveryAnti-HER2 AntibodyTrastuzumabNatural Killer CellsNK Cells3D ModelJIMT-1 Breast Carcinoma CellsGFP96-well PlateAgarose CoatingSpheroid FormationPluronic F-127 CoatingHigh-content Screening

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