A fast and simple fluorometric method to detect cell death in 3D intestinal organoids

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2019
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Schweinlin, Matthias
Metzger, Marco
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Published
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BioTechniques. 2019, 67(1), pp. 23-28. ISSN 0736-6205. eISSN 1940-9818. Available under: doi: 10.2144/btn-2019-0023
Zusammenfassung

Organoids recapitulate the (patho)physiological processes in certain tissues and organs closer than classical cell lines. Therefore, organoid technology offers great potentials in drug development and testing, and personalized medicine. In particular, organoids can be used to study and predict drug-induced toxicity in certain tissues. However, until today few methods had been reported to analyze cell death in 3D-microtissues in a quantitative manner. Here, we describe a novel fluorometric method for the quantitative measurement of specific organoid cell death. Organoids are stained simultaneously with the cell impermeable nuclear dye propidium iodide and cell permeable Hoechst33342. While Hoechst allows in-well normalization to cell numbers, propidium iodide detects relative proportion of dead cells independent of hydrogel. Measurement and analysis time, as well as usability are drastically improved in comparison to other established methods. Parallel multiplexing of our method with established assays measuring mitochondrial activity further enhances its applicability in personalized medicine and drug discovery.

Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
570 Biowissenschaften, Biologie
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cell death, DNA dyes, fluorescence, intestine, organoids, personalized medicine
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Zitieren
ISO 690BODE, Konstantin J., Stefanie MUELLER, Matthias SCHWEINLIN, Marco METZGER, Thomas BRUNNER, 2019. A fast and simple fluorometric method to detect cell death in 3D intestinal organoids. In: BioTechniques. 2019, 67(1), pp. 23-28. ISSN 0736-6205. eISSN 1940-9818. Available under: doi: 10.2144/btn-2019-0023
BibTex
@article{Bode2019-07simpl-46267,
  year={2019},
  doi={10.2144/btn-2019-0023},
  title={A fast and simple fluorometric method to detect cell death in 3D intestinal organoids},
  number={1},
  volume={67},
  issn={0736-6205},
  journal={BioTechniques},
  pages={23--28},
  author={Bode, Konstantin J. and Mueller, Stefanie and Schweinlin, Matthias and Metzger, Marco and Brunner, Thomas}
}
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