Naslov
Mortality curves of individual yeast cells during chronological ageing

Autori
Štafa, Anamarija ; Ni, Ming ; Xu Luping ; Chen, Yong ; Lindner, Ariel.B. ; Taddei, Francois

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Tor, caloric restriction and anabolism in ageing / - , 2010

Skup
Tor, caloric restriction and anabolism in ageing

Mjesto i datum
Split, Hrvatska, 09.2010

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
aging; yeast; microfluidics

Sažetak
The yeast Saccharomyces cerevisiae is the simplest and best characterized eukaryotic organism, widely accepted as a research model for many eukaryotic cell processes. We studied the properties and the dynamics of yeast ageing and death by measuring its chronological life span (CLS), the time a cell can survive in the stationary phase. In the past, this method was typically applied to yeast auxotroph strains in bulk cultures, which potentially introduces multiple confounding effects and decreases the accuracy the CLS measurement. Auxotroph strains are unable to synthesize one or several amino acids and/or nucleic bases and it has been shown by others that they die in short period when exhausted for these particular nutrients and not glucose. In classical CLS experiment, the pH of the media changes over time due to accumulation of acetic acid, while the dead cells lyse and release additional nutrients into the environment. We have indeed found that when grown in batch, number of prototroph cells in the chronologically ageing culture fluctuates and hardly decreases over a two-week period, much longer than previously observed in studies using auxotroph strains. In order to counter these effects, we have developed a microfluidics platform combined with time-lapse fluorescence microscopy, offering single cell resolution and real-time detection of cell death. Most importantly, the continuous flow of fresh, buffered media lacking a carbon source, through our microfluidics chip creates a constant, well-controlled growth environment that can be sustained for weeks, assuring lack of cross-talk between cells. Our initial results show that the maximum life span of prototroph strain in microfluidic platform is on the order of 22 days. Additionally, we observed classical Gompertz ageing curve, characterized by the initial exponential increase in the death rate, followed by a plateau. In the future, our microfluidics platform will allow more precise determination of death rate in different yeast strains as well as better understanding of many complex processes implicated in S. cerevisiae ageing and death.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Biotehnologija

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