Trehalose: A non-permeable cryoprotectant for direct freezing of early stage murine embryos

Krag, K.T.; Koehler, I-M.; Wright, Raymond W.

doi:10.1016/0093-691x(85)90106-2

Theriogenology

1985

DOI: 10.1016/0093-691x(85)90106-2

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Trehalose: A non-permeable cryoprotectant for direct freezing of early stage murine embryos

K.T. Krag¹,

I-M. Koehler²,

Raymond W. Wright³

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Cited by 9 publications

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“…Ethylene glycol is reported as the most effective agent of the glycol derivatives tested [8,9]. Also, ethylene glycol is the least toxic when it is compared with glycerol and propylene glycol in spite of the higher molar lose is effective as a cryoprotectant for direct freezing of 8-to 16-cell embryos [15]. According to some reports [16] trehalose has the highest hydration ability of all osmotic buffers and it can effectively lower the mobility of water molecules hydrogen-bonded with disaccharides.…”

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confidence: 99%

The Efficiency of Ethylene Glycol, Trehalose and Polyvinylpyrrolidone for Successful Vitrification of IVF Bovine Embryos.

Saha

Otoi²,

Suzuki

1996

Journal of Reproduction and Development

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In Experiment I, a suitable combination of a vitrification solution (VS) and a rehydration solution (RS) was investigated for efficient cryopreservation of in vitro fertilized (IVF) bovine blastocysts on day 7. The variable of the VS is the concentration of polyvinylpyrrolidone (PVP) and that of the RS is the concentration of trehalose. Embryo development of 79% and hatching of 73% were obtained with VS 4 , 40% ethylene glycol (EG) + 11.3% trehalose + 12% PVP and RS 1 , 0% trehalose combination. In Experiment II, IVF bovine embryos on days 7, 8 and 9 were vitrified in VS 4 and rehydrated in RS 1. The highest proportions of embryos reexpanded (84%) and hatched (68%) during culture when the age was day 7. In Experiment III, day 7 embryos which had been vitrified in VS 4 and warmed in 30 C waterbath were kept in situ inside the straw at 5 C or 20 C for 5 min before rehydration in RS 1. Significantly higher proportions of embryos developed in culture when they were kept at 5 C than at 20 C. Furthermore, 3 out of 5 recipients in which vitrified embryos had been directly transferred gave birth to normal calves. Therefore, day 7 IVF bovine blastocysts can be successfully vitrified in VS 4 and can be directly transferred into an isotonic environment.

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mentioning

confidence: 99%

The Efficiency of Ethylene Glycol, Trehalose and Polyvinylpyrrolidone for Successful Vitrification of IVF Bovine Embryos.

Saha

Otoi²,

Suzuki

1996

Journal of Reproduction and Development

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Effect of equilibration period on the viability of frozen‐thawed mouse morulae after rapid freezing

Takahashi

Kanagawa

1990

Molecular Reproduction Devel

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Mouse morulae were frozen with 1.5-4.0 M glycerol + 0.25 M lactose solution by direct plunging into liquid nitrogen vapor 0.5-30 min after equilibration at room temperature. After thawing, embryos were cultured in vitro, and the highest survival rates were obtained after exposure for 3 min at 3.0 and 4.0 M and for 5 min at 1.5 and 2.0 M glycerol levels. Significant reductions in the survival rates (P less than 0.05) were observed when equilibration periods were extended for 3-5 min at 3.0 and 4.0 M and for 5-10 min at 1.5 and 2.0 M glycerol levels. These results clearly demonstrate that the equilibration time of embryos in glycerol-lactose mixture is one of the most important factors in the present rapid freezing conditions. To clarify the factors that lower embryo viability after prolonged equilibration, we performed further experiments on the effects of exposure to glycerol-lactose mixture on the developmental potential of embryos without freezing and on the volume changes of embryos during the exposure to glycerol solution with or without lactose. It was suggested that the detrimental effects of prolonged equilibration are due not only to the toxicity and osmotic injury of higher concentrations of cryoprotectant solution but also to the influx of water into embryonic cells caused by the hypotonic salt concentration of the extracellular (freezing) solution.

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The effect of sucrose and trehalose on viability of one- and two-cell rabbit embryos

et al. 1990

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Trehalose: A non-permeable cryoprotectant for direct freezing of early stage murine embryos

Cited by 9 publications

References 0 publications

The Efficiency of Ethylene Glycol, Trehalose and Polyvinylpyrrolidone for Successful Vitrification of IVF Bovine Embryos.

The Efficiency of Ethylene Glycol, Trehalose and Polyvinylpyrrolidone for Successful Vitrification of IVF Bovine Embryos.

Effect of equilibration period on the viability of frozen‐thawed mouse morulae after rapid freezing

The effect of sucrose and trehalose on viability of one- and two-cell rabbit embryos

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