Online citations, reference lists, and bibliographies.
Please confirm you are human
(Sign Up for free to never see this)
← Back to Search

A Flow-cytometric Method For The Separation And Quantitation Of Normal And Apoptotic Thymocytes.

X. Sun, R. T. Snowden, D. Skilleter, D. Dinsdale, M. Ormerod, G. Cohen
Published 1992 · Biology, Medicine

Save to my Library
Download PDF
Analyze on Scholarcy
Using flow cytometry, we describe a method for separating and quantifying normal and apoptotic thymocytes. Apoptosis was induced in isolated thymocytes from immature rats by treatment with the glucocorticoid dexamethasone or the antitumor agent etoposide. Subsequent incubation with the vital bisbenzimidazole dye Hoechst 33342 and the DNA intercalating agent propidium iodide enabled three distinct populations of cells to be identified and sorted by flow cytometry. Dead cells fluoresced red due to propidium iodide whereas normal and apoptotic cells fluoresced blue due to Hoechst 33342. Apoptotic cells were distinguished from normal thymocytes both by their higher intensity of blue fluorescence and by their smaller size as determined by a reduction in forward light scatter. The larger cells, with low blue fluorescence, showed normal thymocyte morphology by electron microscopy and the absence of any DNA fragmentation as measured by agarose gel electrophoresis. In contrast, the smaller cells showed both the morphological characteristics of apoptosis and extensive internucleosomal fragmentation of DNA to multiples of approximately 180 bp. Using this method, a time-dependent induction of apoptosis by dexamethasone, which was inhibited by cycloheximide, actinomycin D, and aurin tricarboxylate, was observed. The method should facilitate mechanistic studies on the induction of apoptosis in thymocytes.
This paper references
Cell death: the significance of apoptosis.
A. Wyllie (1980)
Evaluation of glucocorticoid‐induced DNA fragmentation in mouse thymocytes by flow cytometry
W. Telford (1991)
Chromatin cleavage in apoptosis: Association with condensed chromatin morphology and dependence on macromolecular synthesis
A. Wyllie (1984)
Effect of drug efflux blockers on vital staining of cellular DNA with Hoechst 33342.
A. Krishan (1987)
Hormone-induced cell death. Purification ad properties of thymocytes undergoing apoptosis after glucocorticoid treatment.
A. Wyllie (1982)
Spectral studies on 33258 Hoechst and related bisbenzimidazole dyes useful for fluorescent detection of deoxyribonucleic acid synthesis.
S. Latt (1976)
Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation
A. Wyllie (1980)
Analysis of glucocorticoid actions on rat thymocyte deoxyribonucleic acid by fluorescence-activated flow cytometry.
M. M. Compton (1988)
Apoptosis: mechanisms and roles in pathology.
M. Arends (1991)
A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.
I. Nicoletti (1991)
Analysis and sorting of living cells according to deoxyribonucleic acid content.
D. Arndt-Jovin (1977)
Topoisomerase II-reactive chemotherapeutic drugs induce apoptosis in thymocytes.
P. Walker (1991)
Analysis and discrimination of necrosis and apoptosis (programmed cell death) by multiparameter flow cytometry.
C. Dive (1992)
Analysis of events associated with cell cycle arrest at G2 phase and cell death induced by cisplatin.
C. Sorenson (1990)
Calcium‐activated DNA fragmentation kills immature thymocytes
D. McConkey (1989)

This paper is referenced by
Effect of UV irradiation on cell surface protease activity and amino acid uptake.
T. Piva (1998)
Mechanism of cell death : early morphological changes and DNA degradation of apoptosis in rat thymocytes
X. Sun (1994)
An interleukin‐1β‐converting enzyme‐like protease is a common mediator of apoptosis in thymocytes
H. Fearnhead (1995)
role of cell-cycle arrest and apoptosis of tubular cells Rapamycin impairs recovery from acute renal failure:
J. S. Koh (2016)
The effect of transendothelial migration on eosinophil function.
H. Yamamoto (2000)
Flow Cytometric Detection and Quantification of Apoptotic Cells
T. McCloskey (2006)
Intrathymic injection of anti‐Fas monoclonal antibody prolongs murine non‐vascularized cardiac allograft survival
A. Djamali (2004)
Flow cytometry as a method for studying effects of stressors on primary rat neurons
H. Behbahani (2005)
Selective in vitro inhibition of Leishmania donovani by a semi-purified fraction of wild mushroom Grifola frondosa.
Sirin Salma Sultana (2018)
Apoptosis in human monocytic THP.1 cells involves several distinct targets of N-tosyl-L-phenylalanyl chloromethyl ketone (TPCK)
Huijun Zhu (1997)
Lysophosphatidic acid: a novel growth and survival factor for renal proximal tubular cells.
J. Levine (1997)
Increased apoptosis, changes in intracellular Ca2+, and functional alterations in lymphocytes and macrophages after in vitro exposure to static magnetic field.
D. Flipo (1998)
Factors Modulating Apoptosis: an in-vitro Study in Swine Granulosa Cells
N. Ponderato (2000)
Formation of large molecular weight fragments of DNA is a key committed step of apoptosis in thymocytes.
G. Cohen (1994)
Acute exposure of cultured neurones to ethanol results in reversible DNA single-strand breaks; whereas chronic exposure causes loss of cell viability.
F. Lamarche (2003)
Inhibition of the Raf-1 kinase by cyclic AMP agonists causes apoptosis of v-abl-transformed cells.
E. Weissinger (1997)
Programmed cell death in the developing heart.
M. V. D. van den Hoff (2000)
Regulation of human eosinophil apoptosis
Grant. Dewson (2001)
Programmed Cell Death in the Apical Ganglion During Larval Metamorphosis of the Marine Mollusc Ilyanassa obsoleta
D. Gifondorwa (2006)
Mechanisms of HIV-associated lymphocyte apoptosis.
A. Badley (2000)
Proteasome inhibitor-induced apoptosis of B-chronic lymphocytic leukaemia cells involves cytochrome c release and caspase activation, accompanied by formation of an ∼700 kDa Apaf-1 containing apoptosome complex
J. Almond (2001)
Changes in nuclear chromatin precede internucleosomal DNA cleavage in the induction of apoptosis by etoposide.
X. Sun (1994)
Simvastatin suppresses homocysteine-induced apoptosis in endothelial cells: roles of caspase-3, cIAP-1 and cIAP-2
Z. Xu (2009)
Caractérisation de la mort des cellules animales cultivées en bioréacteur
Sandra Carmaux (2008)
The regulation of mesangial cell apoptosis by inflammatory macrophages
J. Duffield (2001)
Formation of high molecular mass DNA fragments is a marker of apoptosis in the human leukaemic cell line, U937.
G. Bicknell (1994)
α-Actinin-4 Is Required for Normal Podocyte Adhesion*
S. V. Dandapani (2007)
An ICE‐like protease is a common mediator of apoptosis induced by diverse stimuli in human monocytic THP.1 cells
H. Zhu (1995)
Osteoblast cell death on methacrylate polymers involves apoptosis.
J. Gough (2001)
Comparative analysis of flow cytometric methods for apoptosis quantitation in murine thymocytes and human peripheral lymphocytes from controls and HIV-infected persons. Evidence for interference by granulocytes and erythrocytes.
H. Lecoeur (1996)
REVIEW ARTICLE Features of Apoptotic Cells Measured by Flow Cytometry
S. Bruno (1992)
Chronology of cellular alterations during 7‐ketocholesterol–induced cell death on A7R5 rat smooth muscle cells: Analysis by time lapse‐video microscopy and conventional fluorescence microscopy
Jean-Marie Zahm (2003)
See more
Semantic Scholar Logo Some data provided by SemanticScholar