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Article

A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids

by 1,2, 1,3, 4,5, 2,3,6,† and 1,3,7,*,†
1
Cell Death Investigation and Therapy Laboratory, Department of Human Structure and Repair, Ghent University, 9000 Ghent, Belgium
2
Tissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Ghent University, 9000 Ghent, Belgium
3
Cancer Research Institute Ghent, 9000 Ghent, Belgium
4
Plasma, Laser Ablation and Surface Modelling Group, University of Antwerp, 2610 Wilrijk, Belgium
5
Center for Oncological Research, University of Antwerp, 2610 Wilrijk, Belgium
6
Tissue Engineering lab, Department of Development and Regeneration, KU Leuven, 8500 Kortrijk, Belgium
7
Department of Pathophysiology, Sechenov First Moscow State Medical University, 119146 Moscow, Russia
*
Author to whom correspondence should be addressed.
These authors shared senior authorship.
Cells 2020, 9(3), 703; https://doi.org/10.3390/cells9030703
Received: 19 February 2020 / Revised: 6 March 2020 / Accepted: 10 March 2020 / Published: 13 March 2020
(This article belongs to the Section Cell Signaling)
The failure of drug efficacy in clinical trials remains a big issue in cancer research. This is largely due to the limitations of two-dimensional (2D) cell cultures, the most used tool in drug screening. Nowadays, three-dimensional (3D) cultures, including spheroids, are acknowledged to be a better model of the in vivo environment, but detailed cell death assays for 3D cultures (including those for ferroptosis) are scarce. In this work, we show that a new cell death analysis method, named 3D Cell Death Assay (3DELTA), can efficiently determine different cell death types including ferroptosis and quantitatively assess cell death in tumour spheroids. Our method uses Sytox dyes as a cell death marker and Triton X-100, which efficiently permeabilizes all cells in spheroids, was used to establish 100% cell death. After optimization of Sytox concentration, Triton X-100 concentration and timing, we showed that the 3DELTA method was able to detect signals from all cells without the need to disaggregate spheroids. Moreover, in this work we demonstrated that 2D experiments cannot be extrapolated to 3D cultures as 3D cultures are less sensitive to cell death induction. In conclusion, 3DELTA is a more cost-effective way to identify and measure cell death type in 3D cultures, including spheroids. View Full-Text
Keywords: ferroptosis; spheroids; 3D cultures; cell death assay; cancer ferroptosis; spheroids; 3D cultures; cell death assay; cancer
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MDPI and ACS Style

Demuynck, R.; Efimova, I.; Lin, A.; Declercq, H.; Krysko, D.V. A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids. Cells 2020, 9, 703. https://doi.org/10.3390/cells9030703

AMA Style

Demuynck R, Efimova I, Lin A, Declercq H, Krysko DV. A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids. Cells. 2020; 9(3):703. https://doi.org/10.3390/cells9030703

Chicago/Turabian Style

Demuynck, Robin, Iuliia Efimova, Abraham Lin, Heidi Declercq, and Dmitri V. Krysko. 2020. "A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids" Cells 9, no. 3: 703. https://doi.org/10.3390/cells9030703

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