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Published on: November 20, 2021
Vitrification and Ultra-Rapid Laser Warming of Yeast Saccharomyces cerevisiae
1Centro de Investigacion Mariña, Universidade de Vigo, Laboratorio de Ecoloxia Costeira (ECOCOST), Departamento de Ecoloxia e Bioloxia Animal, Vigo, Spain. eparedes@uvigo.es.
Background:
As fundamental model organisms, yeasts have been used for the study and understanding of cryopreservation and freezing damage mechanisms, in particular Saccharomyces cerevisiae.
Objective:
As cryopreservation success requires optimization of the cooling and warming rates, the objective was to test how ultra-rapid warming could improve yeast cell cryopreservation.
Materials And Methods:
S. cerevisiae cells were exposed to concentrations of vitrification solutions containing a combination of permeating and non-permeating cryoprotectants (EAFS) and to a simple 1 M sucrose solution prior to vitrification (cooling rate 69,000°C min-1). Cells were then warmed ultra-rapidly with a laser (warming rate 107°C min-1).
Results:
When using a vitrification solution (0.33xEAFS) survival was 80 ± 16%. When using only a non-permeating solute (1 M sucrose) for cryoprotection, the results were slightly lower, viz. 61 ± 26 %.
Conclusion:
These results add information to the study of the effect of numerous cooling and warming rates for baker´s yeast cryopreservation and provide further examples of the application of vitrification and ultra-fast laser warming. Ultra-rapid warming seems to be applicable to a wide range of cells and tissues from diverse species.
