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Cell death is a fundamental aspect of the life cycle
of the eukaryotic cell. Homeostasis implies a balance between cell proliferation
and cell death, and this balance is essential for the development and
maintenance of multicellular organisms. Dysregulation of either cell death or
proliferation leads to a wide range of pathologies including cancer,
neurodegenerative disease, and aberrant embryogenesis. During the 1990’s
research in the field of cell death grew exponentially, and fundamental aspects
regarding the molecular nature and
physiological
regulation of the dying process were elucidated. By
the end of the decade, t was recognized that cell death occurs through a network
of mechanisms. These mechanisms act in series or in parallel, each of which can
be variably expressed in different eukaryotic species and within the same
species, among distinct cell types. By the early 2000’s assays to analyze cell
death had become essential research and diagnostics tools in laboratories
worldwide.
Apoptosis Concepts and Cell Death
Assays The
main goals of the cell death field is to determine the mechanisms and regulation
of cell death, whether it occurs in prokaryotes, eukaryotes, singlecelled or
multicellular organisms, animals or plants. Traditionally, many cell death
assays have been referred to as apoptosis assays. However, apoptosis was
originally defined purely on morphological terms in a now classic paper by Kerr
et al. in 1972 to designate a morphologically distinct form of cell death
associated with normal physiology. Apoptosis was distinguished from necrosis,
which is associated with acute injury to cells. Apoptosis is characterized by
nuclear chromatin condensation, cell shrinking, dilated endo-plasmic reticulum,
and membrane blebbing.
As the cell death field matured, focus shifted away
from describing the morphological aspects of cell death to assessing the
underlying biology. As a result, today’s assay methodologies are based on
detecting specific biological aspects of the cell death process including
caspase activation, protein cleavage, mitochondrial functioning, loss of
phospholipid membrane asymmetry, and DNA fragmentation.
Many different
models of cell death have emerged including necrosis, apoptosis, anoikis,
caspase-independent apoptosis, autophagy, endoplasmic reticulum stress,
Wallerian degeneration, excitotoxicity, erythropoiesis, platelets, cornification
and lens (reviewed in Melino et al. 2005). However, apoptosis continues to be a
popular, although loosely defined term that is used differently by different
investigators to measure and describe cell death. In general, anything that
looks like apoptosis has been called apoptosis which has contributed to the
complexity of the cell death literature (Vaux et al. 1999). Researchers should
keep in mind that cell death assays detect a biological aspect or mechanism of
cell death versus being exclusive to a particular type of cell death such as
apoptosis or necrosis.
IMGENEX offers a wide range of products and assays
for detecting and analyzing cell death. Collectively, these products encompass a
variety of techniques including flow cytometry, western blot analysis,
immunohistochemisty, fluorescence microscopy, and ELISA. As cell death occurs
through different mechanisms, which may be present in various combinations, the
Nomenclature Committee on Cell Death (NCCD) encourages investigators studying
cell death to use more than one assay whenever possible (Kroemer et al.
2005).
