EMD Millipore eBook - 9

Analyzing Cell Death * So Many Ways to Die

From Within: Intrinsic Induction of Apoptosis
Mitochondrial changes are highly sensitive indicators of cell health and stress.
Cellular energy produced during mitochondrial respiration is stored as an
electrochemical gradient across the mitochondrial membrane, and this accumulation of energy in healthy cells creates a mitochondrial trans-membrane potential
(Ψm) that enables the cell to drive the synthesis of ATP. Loss of the mitochondrial
inner transmembrane potential is often associated with the early stages of apoptosis. Collapse of this potential is believed to coincide with the opening of the
mitochondrial permeability transition pores, leading to the release of cytochrome
C into the cytosol, which then triggers the downstream events in the apoptotic
cascade. Mitochondrial membrane potential changes have been implicated in
apoptosis, necrotic cell death, and caspase-independent cell death processes.
Depolarization of the inner mitochondrial membrane potential is thus a reliable
indicator of mitochondrial dysfunction and cellular health, which has become
increasingly important in the study of apoptosis, drug toxicity and multiple
disease states. The Muse® MitoPotential Assay utilizes the cationic, lipophilic
MitoPotential dye, to detect changes in the mitochondrial membrane
potential (ΔΨm), and 7-AAD as an indicator of cell death.
Quantitative cell analysis: The means to "the end"
Reliable methods for routine analysis of cell health and cell death are critical
for bioprocess scientists in industry, academic investigators, and biomedical
researchers in the basic science, translational, and clinical settings. Furthermore,
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modern research demands data that is both information-rich and that can be
tested for statistical significance. Simplified instruments such as the Muse® Cell
Analyzer that analyze thousands of cells per sample to return powerful cell health
data from optimized assays and reagents represent a new frontier in our ability to
perform postmortem analyses of cell death that yield insights into cell health. n

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Fatokun AA, Dawson VL, and Dawson TM. Parthanatos: mitochondrial-linked
mechanisms and therapeutic opportunities. Br J Pharmacol. 2014: 171: 2000-2016.
Galluzzi L, Kroemer G, et al. Molecular definition of cell death subroutines:
recommendations of the Nomenclature Committee on Cell Death 2012.
Cell Death Differ. 2012 Jan: 19(1): 107-20.
Green DR and Levine B. To be or not to be? How selective autophagy and cell death
govern cell fate. Cell. 2014 March: 157(1): 65-75.
Ouyang L, Shi Z, Zhao S, Wang FT, Zhou TT, Liu B, Bao JK. Programmed cell death
pathways in cancer: a review of apoptosis, autophagy, and programmed necrosis.
Cell Prolif. 2012: 45: 487-498.
Wlodkowic D, Skommer J, Darzynkiewicz Z. Cytometry of apoptosis.
Historical perspective and new advances. Exp Oncol. 2012: 34(3): 255-262.

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