Quantifying yeast chronological life span by outgrowth of aged cells.
The budding yeast Saccharomyces cerevisiae has proven to be an important model organism in the field of aging research. The replicative and chronological life spans are two established paradigms used to study aging in yeast. Replicative aging is defined as the number of daughter cells a single yeast mother cell produces before senescence; chronological aging is defined by the length of time cells can survive in a non-dividing, quiescence-like state. We have developed a high-throughput method for quantitative measurement of chronological life span. This method involves aging the cells in a defined medium under agitation and at constant temperature. At each age-point, a sub-population of cells is removed from the aging culture and inoculated into rich growth medium. A high-resolution growth curve is then obtained for this sub-population of aged cells using a Bioscreen C MBR machine. An algorithm is then applied to determine the relative proportion of viable cells in each sub-population based on the growth kinetics at each age-point. This method requires substantially less time and resources compared to other chronological lifespan assays while maintaining reproducibility and precision. The high-throughput nature of this assay should allow for large-scale genetic and chemical screens to identify novel longevity modifiers for further testing in more complex organisms.
Department of Pathology, University of Washington, Seattle, WA 98195-7470, USA.
Christopher
C
Murakami
Matt
M
Kaeberlein
D048708:Cell Growth Processes / Q000502:physiology; D016922:Cellular Senescence / Q000502:physiology*; D009172:Mycology / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000502:physiology*