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A 3D cell death assay to quantitatively determine ferroptosis in spheroids

Robin Demuynck (UGent) , Iuliia Efimova (UGent) , Abraham Lin, Heidi Declercq (UGent) and Dmitri Krysko (UGent)
(2020) CELLS. 9(3).
Author
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Abstract
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.</jats:p>
Keywords
ferroptosis, spheroids, 3D cultures, cell death assay, cancer, CANCER, FLUORESCENCE, RESISTANCE, APOPTOSIS, AUTOPHAGY, NECROSIS, MODELS, 2D

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MLA
Demuynck, Robin, et al. “A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids.” CELLS, vol. 9, no. 3, 2020, doi:10.3390/cells9030703.
APA
Demuynck, R., Efimova, I., Lin, A., Declercq, H., & Krysko, D. (2020). A 3D cell death assay to quantitatively determine ferroptosis in spheroids. CELLS, 9(3). https://doi.org/10.3390/cells9030703
Chicago author-date
Demuynck, Robin, Iuliia Efimova, Abraham Lin, Heidi Declercq, and Dmitri Krysko. 2020. “A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids.” CELLS 9 (3). https://doi.org/10.3390/cells9030703.
Chicago author-date (all authors)
Demuynck, Robin, Iuliia Efimova, Abraham Lin, Heidi Declercq, and Dmitri Krysko. 2020. “A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids.” CELLS 9 (3). doi:10.3390/cells9030703.
Vancouver
1.
Demuynck R, Efimova I, Lin A, Declercq H, Krysko D. A 3D cell death assay to quantitatively determine ferroptosis in spheroids. CELLS. 2020;9(3).
IEEE
[1]
R. Demuynck, I. Efimova, A. Lin, H. Declercq, and D. Krysko, “A 3D cell death assay to quantitatively determine ferroptosis in spheroids,” CELLS, vol. 9, no. 3, 2020.
@article{8654224,
  abstract     = {{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.</jats:p>}},
  articleno    = {{703}},
  author       = {{Demuynck, Robin and Efimova, Iuliia and Lin, Abraham and Declercq, Heidi and Krysko, Dmitri}},
  issn         = {{2073-4409}},
  journal      = {{CELLS}},
  keywords     = {{ferroptosis,spheroids,3D cultures,cell death assay,cancer,CANCER,FLUORESCENCE,RESISTANCE,APOPTOSIS,AUTOPHAGY,NECROSIS,MODELS,2D}},
  language     = {{eng}},
  number       = {{3}},
  pages        = {{13}},
  title        = {{A 3D cell death assay to quantitatively determine ferroptosis in spheroids}},
  url          = {{http://dx.doi.org/10.3390/cells9030703}},
  volume       = {{9}},
  year         = {{2020}},
}

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