1. Isolation of quiescent and nonquiescent cells from yeast stationary-phase cultures
- Author
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Frank Madeo, Stephanie W. Ruby, Osorio Meirelles, Chris Allen, Jason E. Jaetao, Margaret Werner-Washburne, Jason A. Thomas, Don Benn, Sabrina Büttner, Marten Veenhuis, Anthony D. Aragon, Groningen Biomolecular Sciences and Biotechnology, and Molecular Cell Biology
- Subjects
STRESS ,Cell division ,Saccharomyces cerevisiae ,Mitosis ,Cell Separation ,Resting Phase, Cell Cycle ,Sensitivity and Specificity ,Article ,Flow cytometry ,SACCHAROMYCES-CEREVISIAE ,SUPEROXIDE ,03 medical and health sciences ,0302 clinical medicine ,Mitotic cell cycle ,EXIT ,medicine ,Centrifugation, Density Gradient ,Research Articles ,Cells, Cultured ,030304 developmental biology ,GENE-EXPRESSION ,0303 health sciences ,Microscopy ,biology ,medicine.diagnostic_test ,IDENTIFICATION ,Cell Cycle ,Cell Biology ,DNA ,Cell cycle ,biology.organism_classification ,Flow Cytometry ,Yeast ,Cell biology ,APOPTOSIS ,Glucose ,Apoptosis ,COLONIES ,Reactive Oxygen Species ,REGULATOR ,030217 neurology & neurosurgery - Abstract
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 G0 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.
- Published
- 2006