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Current status of vitrification of embryos and oocytes in domestic animals: Ethylene glycol as an emerging cryoprotectant of choice
1998
Bautista, J.A.N. (Hokkaido Univ., Sapporo (Japan)) | Kanagawa, H.
The cryopreservation of mammalian embryos has become an integral part of method s to control animal reproduction. Numerous vitrification solutions have been formulated with ethylene glycol in combination with macromolecules, sugars and other cryoprotective agents. These indicate that a study of ethylene glycol as a cryoprotectant of choice in vitrification studies would be promising. To understand the cryobiology of ethylene glycol, several factors have to be studied. These are : cryoprotectant toxicity, osmotic stress and temperature at exposure. Understanding these factors could lead to the formulation of vitrification protocols that would lead to higher viability rates after cooling. First, ethylene glycol must be used as the sole cryoprotectant in a solution without macromolecules and sugars. Second, partial dehydration and permeation prior to cooling to subzero temperatures must be studied to achieve accurate exposure and a one-step dilution method. Third, the toxic effects of ethylene glycol must be overcome without sacrificing its vitrification properties by combining step-wise exposure at appropriate temperatures, low concentration and decreased volume. Fourth, the long-term effects of ethylene glycol on exposed or vitrified embryos must be determined. Lastly, the influence of culture on the viability of vitrified embryos must be studied to improve viability rates after warming
Show more [+] Less [-]In vitro viability of mouse oocytes vitrified in an ethylene glycol-based solution
1998
Bautista, J.A.N. (Hokkaido Univ., Sapporo (Japan)) | Pena, E.C.D. | Katagiri, S. | Takahashi, Y. | Kanagawa, H.
Ovulated mouse oocytes denuded of their cumulus cells, were vitrified in a solution containing 7 M ethylene glycol as the sole cryoprotectant using one or two steps of exposure before vitrification and were diluted in 1 M sucrose solution in 5 or 10 min after warming. The results proved that the viability of oocytes are detrimentally affected by exposure to the vitrification solution even without vitrification. At 5 min dilution time, the two-step exposure was superior to the one-step in terms of the post-warming recovery rate of vitrified oocytes with normal morphology and their subsequent development to the blastocyst stage (p0.001) after fertilization in vitro. At 10 min dilution time, no significant difference between one or two-step exposure was found. The effect of the addition of 0.5 M sucrose to the vitrification solution was also determined and did not result in a significant improvement in the viability of oocytes vitrified in one-step and diluted for 10 min. In conclusion, the results in this study indicate that oocytes can be vitrified with 7 M ethylene glycol as the sole cryoprotectant in the vitrification solution, and that the recovery of normal oocytes after one-step exposure in the vitrification solution can be improved by 10 min dilution time. However, the improvement in the recovery rate of oocytes with normal morphology and their subsequent developmental in vitro was not improved by the addition of 0.5 M sucrose to the vitrification solution
Show more [+] Less [-]Фенотипические особенности ремонтных телочек, полученных методом трансплантации замороженно-оттаянных эмбрионов
2009
Minina, N.G., Grodno State Agrarian Univ. (Belarus)
The study has stated that heifer replacements, from the date of birth to 18 months, got through transplantation of frozen-defrosted embryos were not inferior to and in many cases superior to the heifers of their age but got through traditional reproduction. Transplant first-calf heifers have a high level of milk productivity which amounts to 7536,10 kilos, with 3,62% of fat and 3,23% of protein. Thus they provide a good selection material for new generation champions to be chosen among them and for being used as mothers of bulls for service
Show more [+] Less [-]Использование биологически активных веществ при созревании ооцитов крупного рогатого скота вне организма
2009
Rakovich, E.D., National Academy of Sciences. Scientific and Practical Center of Animal Breeding (Belarus)
The problem of creation of improved conditions for cattle oocyte maturation in vitro was studied in the Republic of Belarus. The objects of study were cattle oocytes and embryos of early preimplantation stages of development obtained in vitro conditions. Research results showed that application of bovine serum in complex with embryonic or fetal serum in the process of oocyte cultivation in vitro conditions made it possible to obtain 35,5-44,8% of breaking down cells and 16,1-17,2 % of embryos at the stage of morula-blastocyte. Entering of 0,02 ng/ml of gonadotropin-releasing hormone (Surfagon) into media for oocyte maturation at the cost of regulation of the processes of growing and development of oocyte-cumulus complexes made it possible to increase of yield of matured till the stage of metaphase II oocytes up to 89,3 %. At the same time, the yield of divided embryos after fertilization increased on 2,5 % in comparison with control indexes and amounted 45,2 %. Research results showed that the synthetic phytohormone epibrassinolide could be used as a biologically active factor in the process of getting early embryos in vitro in concentration of 2 x 10E-7 – 2 x 10E-9 mole/l. It made it possible to get 46,4-55,0 % of divided cells and 14,2-16,2% preimplantation embryos
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