Document Detail

Supravital exposure to propidium iodide identifies apoptotic cells in the absence of nucleosomal DNA fragmentation.
MedLine Citation:
PMID:  8900473     Owner:  NLM     Status:  MEDLINE    
By flow cytometry, we have quantitatively evaluated HL-60, MOLT-4, and P815 apoptotic cells induced with camptothecin, staurosporine, and hyperthermia, respectively. Apoptosis was measured by two different flow cytometry techniques, using propidium iodide (PI) uptake in permeabilized and nonpermeabilized cells. Apoptosis also has been analyzed by electron microscopy and DNA cleavage by in situ nick translation and DNA gel electrophoresis. We demonstrate that supravital exposure to propidium iodide without prior permeabilization identifies apoptotic cells, and clearly distinguishes them from necrotic cells in all the cases examined. This capability is independent of the nucleosomal (180-200 bp) fragmentation of DNA (which does not take place in MOLT-4 cells), which is the basis for detection of apoptosis both as a "ladder" by DNA gel electrophoresis and as a hypodiploid peak by flow cytometry. Therefore, alterations in membrane permeability, on which PI uptake in living cells is based, allow distinction of apoptotic cells from necrotic and living cells independently of the heterogeneous biochemical patterns involved in programmed cell death, which may or may not lead to DNA oligonucleosomal fragmentation.
L Zamai; E Falcieri; G Marhefka; M Vitale
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry     Volume:  23     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  1996 Apr 
Date Detail:
Created Date:  1996-12-23     Completed Date:  1996-12-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  303-11     Citation Subset:  IM    
Istituto di Anatomia Umana, Università di Bologna, Italy.
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MeSH Terms
Apoptosis / physiology*
Coloring Agents / pharmacology*
DNA Damage / physiology*
Flow Cytometry / methods*
HL-60 Cells
Propidium / pharmacology*
Tumor Cells, Cultured
Reg. No./Substance:
0/Coloring Agents; 0/Nucleosomes; 36015-30-2/Propidium

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