Format

Send to

Choose Destination
Yeast. 2017 Aug;34(8):343-355. doi: 10.1002/yea.3237. Epub 2017 Jun 6.

A multiplex culture system for the long-term growth of fission yeast cells.

Author information

1
SyntheCell Team, Institute of Genetics and Development of Rennes, CNRS UMR 6290, Rennes, France.
2
Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
3
Ecole National Supérieure d'Ingénieurs de Caen, UMR 6072 - GREYC, Caen, France.

Abstract

Maintenance of long-term cultures of yeast cells is central to a broad range of investigations, from metabolic studies to laboratory evolution assays. However, repeated dilutions of batch cultures lead to variations in medium composition, with implications for cell physiology. In Saccharomyces cerevisiae, powerful miniaturized chemostat setups, or ministat arrays, have been shown to allow for constant dilution of multiple independent cultures. Here we set out to adapt these arrays for continuous culture of a morphologically and physiologically distinct yeast, the fission yeast Schizosaccharomyces pombe, with the goal of maintaining constant population density over time. First, we demonstrated that the original ministats are incompatible with growing fission yeast for more than a few generations, prompting us to modify different aspects of the system design. Next, we identified critical parameters for sustaining unbiased vegetative growth in these conditions. This requires deletion of the gsf2 flocculin-encoding gene, along with addition of galactose to the medium and lowering of the culture temperature. Importantly, we improved the flexibility of the ministats by developing a piezo-pump module for the independent regulation of the dilution rate of each culture. This made it possible to easily grow strains that have different generation times in the same assay. Our system therefore allows for maintaining multiple fission yeast cultures in exponential growth, adapting the dilution of each culture over time to keep constant population density for hundreds of generations. These multiplex culture systems open the door to a new range of long-term experiments using this model organism. © 2017 The Authors. Yeast published by John Wiley & Sons, Ltd.

KEYWORDS:

Schizosaccharomyces; chemostats; continuous culture; experimental evolution; turbidostats

PMID:
28426144
PMCID:
PMC5542872
DOI:
10.1002/yea.3237
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for PubMed Central
Loading ...
Support Center