Document Detail

Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation.
MedLine Citation:
PMID:  18388949     Owner:  NLM     Status:  MEDLINE    
Synchronization of cells and nuclei is a powerful technique for the exact study of regulatory mechanisms and for understanding cell cycle events. Counterflow centrifugal elutriation is a biophysical cell separation technique in which cell size and sedimentation density differences of living cells are exploited to isolate subpopulations in various stages of cell cycle. Here, a protocol is described for the separation of phase-enriched subpopulations from exponentially growing Chinese hamster ovary cells at high-resolution power of elutriation. The efficiency of elutriation is confirmed by measuring the DNA content fluorimetrically and by flow cytometry. The resolution power of elutriation is demonstrated by the ability to fractionate nuclei of murine pre-B cells. The installation and elutriation by collecting 16-30 synchronized fractions, including particle size analysis, can be achieved in 4-5 h.
Gaspar Banfalvi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  3     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2008  
Date Detail:
Created Date:  2008-04-04     Completed Date:  2008-10-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  663-73     Citation Subset:  IM    
Department of Microbial Biotechnology and Cell Biology, University of Debrecen, Debrecen 4010, Hungary.
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MeSH Terms
Cell Cycle / physiology*
Cell Division
Cell Line
Cell Nucleus / physiology*
Cell Separation / methods
Centrifugation, Density Gradient / methods*
Epithelial Cells / cytology*

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