Ultrarapid freezing of rat embryos with rapid dilution of permeable cryoprotectants. 1997

V V Isachenko, and E F Isachenko, and F I Ostashko, and V I Grishchenko
Department of Animal Reproduction, Institute for Animal Science, Kharkov, Ukraine.

The purpose of this study was to determine the feasibility of ultrarapid freezing of rat morulae with rapid postthaw dilution of permeable cryoprotectants in isotonic culture medium. Four experiments were carried out. Experiment 1 examined the possibility of using vitrification with postthaw dilution of permeable cryoprotectants in an isotonic solution. Embryos were exposed first to 10% glycerol + 20% propylene glycol and then to the final vitrification solution which contained 25% glycerol + 25% propylene glycol. Embryo survival was very low when the subsequent dilution was in a solution that did not contain sucrose. In Experiment 2. three mixtures were tested: 15% glycerol + 15% ethylene glycol + 0.7 M sucrose, 15% glycerol + 15% propylene glycol + 0.7 M sucrose, and 30% glycerol + 0.7 M sucrose. The third mixture, which contained only glycerol and sucrose, produced the best results with 88% embryo survival. In Experiment 3, the embryos were frozen in 30% glycerol plus 0.7 M sucrose and in addition were exposed to 1 M sucrose for 7 min following thawing. The survival rate was 85% with the sucrose dilution step, 91% when dilution was in isotonic medium, and 95% in controls not exposed to the cryoprotective mixture. Experiment 4 examined the effect of the time and temperature of exposure of the embryos to 30% glycerol + 0.7 M sucrose. The highest rates of embryo development followed exposure at 4 degrees C for 2-3 min (95-84%) or at 24 degrees C for 0.5-3.0 min (90-88%). These results indicate that it is possible to develop a method for the ultrarapid freezing of mammalian embryos that does not require dilution of permeable cryoprotectants in a hypertonic sucrose solution.

UI MeSH Term Description Entries
D008297 Male Males
D009028 Morula An early embryo that is a compact mass of about 16 BLASTOMERES. It resembles a cluster of mulberries with two types of cells, outer cells and inner cells. Morula is the stage before BLASTULA in non-mammalian animals or a BLASTOCYST in mammals. Morulas
D010539 Permeability Property of membranes and other structures to permit passage of light, heat, gases, liquids, metabolites, and mineral ions. Permeabilities
D003451 Cryoprotective Agents Substances that provide protection against the harmful effects of freezing temperatures. Cryoprotective Agent,Cryoprotective Effect,Cryoprotective Effects,Agent, Cryoprotective,Agents, Cryoprotective,Effect, Cryoprotective,Effects, Cryoprotective
D004622 Embryo, Mammalian The entity of a developing mammal (MAMMALS), generally from the cleavage of a ZYGOTE to the end of embryonic differentiation of basic structures. For the human embryo, this represents the first two months of intrauterine development preceding the stages of the FETUS. Embryonic Structures, Mammalian,Mammalian Embryo,Mammalian Embryo Structures,Mammalian Embryonic Structures,Embryo Structure, Mammalian,Embryo Structures, Mammalian,Embryonic Structure, Mammalian,Embryos, Mammalian,Mammalian Embryo Structure,Mammalian Embryonic Structure,Mammalian Embryos,Structure, Mammalian Embryo,Structure, Mammalian Embryonic,Structures, Mammalian Embryo,Structures, Mammalian Embryonic
D005069 Evaluation Studies as Topic Works about studies that determine the effectiveness or value of processes, personnel, and equipment, or the material on conducting such studies. Critique,Evaluation Indexes,Evaluation Methodology,Evaluation Report,Evaluation Research,Methodology, Evaluation,Pre-Post Tests,Qualitative Evaluation,Quantitative Evaluation,Theoretical Effectiveness,Use-Effectiveness,Critiques,Effectiveness, Theoretical,Evaluation Methodologies,Evaluation Reports,Evaluation, Qualitative,Evaluation, Quantitative,Evaluations, Qualitative,Evaluations, Quantitative,Indexes, Evaluation,Methodologies, Evaluation,Pre Post Tests,Pre-Post Test,Qualitative Evaluations,Quantitative Evaluations,Report, Evaluation,Reports, Evaluation,Research, Evaluation,Test, Pre-Post,Tests, Pre-Post,Use Effectiveness
D005260 Female Females
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor

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