Document Detail


A rapid and optimization-free procedure allows the in vivo detection of subtle cell cycle and ploidy alterations in tissues by flow cytometry.
MedLine Citation:
PMID:  20928939     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cell cycle alterations are fundamental to many physiological processes but their detection has proven difficult when cells are in the context of a tissue structure. Here we describe an easy, rapid and optimization-free procedure for obtaining high resolution cell cycle profiles from nearly all tissue types derived from mouse, human and sheep. Using a standardized and non-enzymatic procedure that is universally suitable for soft, solid and epithelial tissues alike, we reproducibly obtain cell cycle profiles of highest quality with half peak coefficients of variation below 2.0. We are able to reduce preparation-derived debris to almost zero and efficiently exclude doublets, but retain multinucleated cells and apoptotic subG1-fragments. Applying this technique, we determine DNA-indices as small as 1.09 in tumor samples containing large necrotic areas and follow ploidy changes within different sections of individual tumors. Moreover, we examine tissue-specific cell cycle arrest and apoptosis as an in vivo stress response caused by radiation of mice. This method significantly improves the quality of DNA content analysis in tissues and extends the spectrum of applications. It allows assessing changes in ploidy, cell cycle distribution and apoptosis/necrosis in vivo and should be instrumental in all research that involves experimental animal models and/or patient biopsies.
Authors:
Christina Heinlein; Wolfgang Deppert; Antony W Braithwaite; Daniel Speidel
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell cycle (Georgetown, Tex.)     Volume:  9     ISSN:  1551-4005     ISO Abbreviation:  Cell Cycle     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-10-06     Completed Date:  2011-01-24     Revised Date:  2011-07-28    
Medline Journal Info:
Nlm Unique ID:  101137841     Medline TA:  Cell Cycle     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3584-90     Citation Subset:  IM    
Affiliation:
Children’s Medical Research Institute, Westmead, Australia.
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MeSH Terms
Descriptor/Qualifier:
Aneuploidy
Animals
Apoptosis
Cell Cycle*
DNA / analysis*
Flow Cytometry / methods*
G1 Phase
Humans
Mice
Mice, Inbred C57BL
Ploidies*
Sheep
Chemical
Reg. No./Substance:
9007-49-2/DNA
Comments/Corrections
Comment In:
Cell Cycle. 2010 Sep 1;9(17):3395-6   [PMID:  20861663 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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