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The use of 7-amino actinomycin D in identifying apoptosis: simplicity of
use and broad spectrum of application compared with other techniques
NJ Philpott, AJ Turner, J Scopes, M Westby, JC Marsh, EC Gordon-Smith, AG Dalgleish and FM Gibson
Division of Haematology, Department of Cellular and Molecular Sciences, St.
George's Hospital Medical School, London, UK.
The detection and quantitation of apoptotic cells is becoming increasingly
important in the investigation of the role of apoptosis in cellular
proliferation and differentiation. The pathogenesis of hematologic
disorders such as aplastic anemia and the development of neoplasia are
believed to involve dysregulation of apoptosis. To quantitate accurately
the proportion of apoptosis cells within different cell types of a
heterogeneous cell population such as blood or bone marrow, a method is
required that combines the analysis of large numbers of cells with
concurrent immunophenotyping of cell surface antigens. In this study, we
have evaluated such a method using the fluorescent DNA binding agent,
7-amino actinomycin D (7AAD), to stain three diverse human cell lines,
induced to undergo apoptosis by three different stimuli. Flow cytometric
analysis defines three populations on the basis of 7AAD fluorescence and
forward light scatter. We have shown by cell sorting and subsequent
morphological assessment and terminal deoxynucleotidyl transferase-mediated
deoxyuridine triphosphate nick end labeling that the populations defined by
7AAD represent live, apoptotic, and late-apoptotic/dead cells. This method
is quick, simple, reproducible, and cheap and will be a valuable tool in
the investigation of the role of apoptosis in normal physiology and in
disease states.
Volume 87,
Issue 6,
pp. 2244-2251,
03/15/1996
Copyright © 1996 by The American Society of Hematology
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