Abstract
Cell death and its recently discovered regulated form ferroptosis are characterized by distinct morphological, electrophysiological, and pharmacological features. In particular ferroptosis can be induced by experimental compounds and clinical drugs (i.e., erastin, sulfasalazine, sorafenib, and artesunate) in various cell types and cancer cells. Pharmacologically, this cell death process can be inhibited by iron chelators and lipid peroxidation inhibitors. Relevance of this specific cell death form has been found in different pathological conditions such as cancer, neurotoxicity, neurodegeneration, and ischemia. Distinguishing cell viability and cell death is essential for experimental and clinical applications and a key component in flow cytometry experiments. Dead cells can compromise the integrity of the data by nonspecific binding of antibodies and dyes. Therefore it is essential that dead cells are robustly and reproducibly identified and characterized by means of cytometry application. Here we describe a procedure to detect and quantify cell death and its specific form ferroptosis based on standard flow cytometry techniques.
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References
Edwards BS, Sklar LA (2015) Flow cytometry: impact on early drug discovery. J Biomol Screen 20(6):689–707
Jaye DL, Bray RA, Gebel HM et al (2012) Translational applications of flow cytometry in clinical practice. J Immunol 188(10):4715–4719
Crosland-Taylor PJ (1953) A device for counting small particles suspended in a fluid through a tube. Nature 171(4340):37–38
Hallermann L, Thom R, Gerhartz H (1964) Electronic differential counting of granulocytes and lymphocytes after intravital fluorochrome staining with acridine organe. Verh Dtsch Ges Inn Med 70:217–219
Fulwyler MJ (1965) Electronic separation of biological cells by volume. Science 150:910–911
Kamentsky LA, Melamed MR, Derman H (1965) Spectrophotometer: new instrument for ultrarapid cell analysis. Science 150:630–631
Wilder ME, Cram LS (1977) Differential fluorochromasia of human lymphocytes as measured by flow cytometry. J Histochem Cytochem 25:888–891
Bendall SC, Nolan GP, Roederer M, Chattopadhyay PK (2012) A deep profiler’s guide to cytometry. Trends Immunol 33:323–332
Bandura DR, Baranov VI, Ornatsky OI, Antonov A, Kinach R, Lou X, Pavlov S et al (2009) Mass cytometry: technique for real time single cell multitarget immunoassay based on inductively coupled plasma time-of-flight mass spectrometry. Anal Chem 81:6813–6822
Perfetto SP, Chattopadhyay PK, Roederer M (2004) Seventeen-colour flow cytometry: unravelling the immune system. Nat Rev Immunol 4:648–655
Sehm T, Fan Z, Ghoochani A, Rauh M et al (2016) Sulfasalazine impacts on ferroptotic cell death and alleviates the tumor microenvironment and glioma-induced brain edema. Oncotarget. doi:10.18632/oncotarget.8651
Sehm T, Rauh M, Wiendieck K, Buchfelder M, Eyüpoglu IY, Savaskan NE (2016) Temozolomide toxicity operates in a xCT/SLC7a11 dependent manner and is fostered by ferroptosis. Oncotarget 7(46):74630–74647. doi: 10.18632/oncotarget.11858.
Chen D, Fan Z, Rauh M, Buchfeller M, Eyupoglu IY, Savaskan N. ATF4 promotes angiogenesis and neuronal cell death and confers ferroptosis in a xCT-dependent manner. Oncotarget 2017 (in press).
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Chen, D., Eyupoglu, I.Y., Savaskan, N. (2017). Ferroptosis and Cell Death Analysis by Flow Cytometry. In: Gilbert, D., Friedrich, O. (eds) Cell Viability Assays. Methods in Molecular Biology, vol 1601. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6960-9_6
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DOI: https://doi.org/10.1007/978-1-4939-6960-9_6
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6960-9
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