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A Method for High-Throughput Quantitative Analysis of Yeast Chronological Life Span

Departments of ¹ Pathology and ² Biochemistry, University of Washington, Seattle.

Address correspondence to Matt Kaeberlein, PhD, Department of Pathology, University of Washington, Box 357470, Seattle, WA 98195-7470. E-mail: kaeber{at}u.washington.edu

Chronological aging in yeast has been studied by maintaining cells in a quiescent-like stationary phase culture and monitoring cell survival over time. The composition of the growth medium can have a profound influence on chronological aging. For example, dietary restriction accomplished by lowering the glucose concentration of the medium significantly increases life span. Here we report a novel high-throughput method for measuring yeast chronological life span by monitoring outgrowth of aging cells using a Bioscreen C MBR machine. We show that this method provides survival data comparable to traditional methods, but with decreased variability. In addition to reducing the glucose concentration, we find that elevated amino acid levels or increased osmolarity of the growth medium is sufficient to increase chronological life span. We also report that life-span extension from dietary restriction does not require any of the five yeast sirtuins (Sir2, Hst1, Hst2, Hst3, or Hst4) either alone or in combination.

Key Words: Longevity—Dietary restriction—Glucose—Amino acids—Osmolarity—Sir2