Isolation of quiescent and nonquiescent cells from yeast stationary-phase cultures

Quiescence is the most common and, arguably, most poorly understood cell cycle state. This is in part because pure populations of quiescent cells are typically difficult to isolate. We report the isolation and characterization of quiescent and nonquiescent cells from stationary-phase (SP) yeast cult...

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Published in:The Journal of cell biology Vol. 174; no. 1; p. 89
Main Authors: Allen, Chris, Büttner, Sabrina, Aragon, Anthony D, Thomas, Jason A, Meirelles, Osorio, Jaetao, Jason E, Benn, Don, Ruby, Stephanie W, Veenhuis, Marten, Madeo, Frank, Werner-Washburne, Margaret
Format: Journal Article
Language:English
Published: United States 03.07.2006
Subjects:
Apoptosis - physiology
Cell Cycle - physiology
Cell Separation - methods
Cells, Cultured
Centrifugation, Density Gradient - methods
Flow Cytometry
Glucose - chemistry
Microscopy - methods
Mitosis
Reactive Oxygen Species - metabolism
Resting Phase, Cell Cycle - physiology
Saccharomyces cerevisiae - cytology
Saccharomyces cerevisiae - isolation & purification
Sensitivity and Specificity
ISSN:0021-9525
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Summary:Quiescence is the most common and, arguably, most poorly understood cell cycle state. This is in part because pure populations of quiescent cells are typically difficult to isolate. We report the isolation and characterization of quiescent and nonquiescent cells from stationary-phase (SP) yeast cultures by density-gradient centrifugation. Quiescent cells are dense, unbudded daughter cells formed after glucose exhaustion. They synchronously reenter the mitotic cell cycle, suggesting that they are in a G(0) state. Nonquiescent cells are less dense, heterogeneous, and composed of replicatively older, asynchronous cells that rapidly lose the ability to reproduce. Microscopic and flow cytometric analysis revealed that nonquiescent cells accumulate more reactive oxygen species than quiescent cells, and over 21 d, about half exhibit signs of apoptosis and necrosis. The ability to isolate both quiescent and nonquiescent yeast cells from SP cultures provides a novel, tractable experimental system for studies of quiescence, chronological and replicative aging, apoptosis, and the cell cycle.
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ISSN:0021-9525
DOI:10.1083/jcb.200604072