138. Survival of common carp Cyprinus Carpio embryos after chorion modification by alcalase and following incubation in vitrifying solutions

Dzyuba, Borys; Boryshpolets, Sergii; Pšenička, Martin; Rodina, Marek; Gela, David; Linhart, O.

doi:10.1016/j.cryobiol.2010.10.142

Cited by 2 publications

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“…We studied the molecular structure of the quenched aqueous mixtures which had been used in the fish embryo cryopreservation experiments described previously [ 9 ]. As is shown in figure 2 , only the pure PD (Samples 1 and 2) and the samples with 23 vol% PD, 17 vol% Met and 20 vol% DMSO mixture in water, samples 4–7, produce diffraction patterns typical of a glass structure.…”

Section: Resultsmentioning

confidence: 99%

“…The cryoprotectant solutions used here were formulated during a previous study of fish (common carp) embryo cryopreservation [ 9 ]. In that work, the possible modification of the chorion of common carp ( Cyprinus carpio ) embryos by alcalase (proteolytic enzyme) treatment to increase the chorion permeability for cryoprotectants was examined.…”

Section: Introductionmentioning

confidence: 99%

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Segregated water observed in a putative fish embryo cryopreservative

et al. 2016

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Development of new cryopreservation strategies has major potential in medicine and agriculture and is critical to the conservation of endangered species that currently cannot be preserved. A critical property of any potential cryopreservative solution is its ability to prevent cell-damaging ice formation during cooling and subsequent heating. This study focuses on the freezing behaviour of promising model cryoprotective solutions. We perform neutron scattering analysis, combined with computer modelling, of the water structure after quench cooling these solutions. It is found that water in this solution forms nano-clusters encapsulated by the surrounding matrix of cryoprotectant solute molecules. We posit that these small volumes inhibit ice formation, because water does not have space for the structural relaxation required to crystallize on the timescale of the cooling process.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Segregated water observed in a putative fish embryo cryopreservative

et al. 2016

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“…The neutron scattering data were obtained on the small angle diffractometer SANDALS [ 8 ] at the ISIS Facility, STFC Rutherford Appleton Laboratory, UK. In the experiment we studied cryo-protectant solutions sample with 23vol% 1,2-propanediol (PD), 17vol% methanol (Met), 20vol% dimethylsulphoxide (DMSO) mixture in water, the sample was formulated during a previous study of fish (Common carp) embryo cryopreservation [ 7 ]. The required chemical components were obtained from Sigma-Aldrich and mixed by volume to the required concentrations.…”

Section: Experimental Methods and Resultsmentioning

confidence: 99%

Strong Isotope Effects on Melting Dynamics and Ice Crystallisation Processes in Cryo Vitrification Solutions

et al. 2015

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The nucleation and growth of crystalline ice during cooling, and further crystallization processes during re-warming are considered to be key processes determining the success of low temperature storage of biological objects, as used in medical, agricultural and nature conservation applications. To avoid these problems a method, termed vitrification, is being developed to inhibit ice formation by use of high concentration of cryoprotectants and ultra-rapid cooling, but this is only successful across a limited number of biological objects and in small volume applications. This study explores physical processes of ice crystal formation in a model cryoprotective solution used previously in trials on vitrification of complex biological systems, to improve our understanding of the process and identify limiting biophysical factors. Here we present results of neutron scattering experiments which show that even if ice crystal formation has been suppressed during quench cooling, the water molecules, mobilised during warming, can crystallise as detectable ice. The crystallisation happens right after melting of the glass phase formed during quench cooling, whilst the sample is still transiting deep cryogenic temperatures. We also observe strong water isotope effects on ice crystallisation processes in the cryoprotectant mixture. In the neutron scattering experiment with a fully protiated water component, we observe ready crystallisation occurring just after the glass melting transition. On the contrary with a fully deuteriated water component, the process of crystallisation is either completely or substantially supressed. This behaviour might be explained by nuclear quantum effects in water. The strong isotope effect, observed here, may play an important role in development of new cryopreservation strategies.

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138. Survival of common carp Cyprinus Carpio embryos after chorion modification by alcalase and following incubation in vitrifying solutions

Cited by 2 publications

References 0 publications

Segregated water observed in a putative fish embryo cryopreservative

Segregated water observed in a putative fish embryo cryopreservative

Strong Isotope Effects on Melting Dynamics and Ice Crystallisation Processes in Cryo Vitrification Solutions

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