CRYOPRESERVATION OF IN VITRO PRODUCED BOVINE EMBRYOS
Keywords:
cattle, IVF, IVC, cryopreservation, vitrificationAbstract
In order to compare the survival rate of bovine embryos frozen by different protocols, oocytes
derived from ovaries in slaughterhouses were used for IVM, IVF and IVC. Expanded blastocysts
were cryopreserved by slow freezing at 0.3 ºC/min, and vitrification was done using ethylene glycol
+ sucrose (ES). After thawing, the embryos were co-cultivated on a monolayer of granulosa cells
and evaluated after 24 hours for re-expansion. At 24, 48, 72 and 96 hours the stages of development
and hatching were determined. The degree of in vitro survival (hatching of 96 hours) of frozen by
slow freezing method embryos at a cooling rate of 0.3 ºC/min (76.44%) was similar to that obtained
for the control group (88.8% without cryopreservation). Vitrification using ES showed survival rates
of 18.9%. In conclusion, in our hands, the slow freezing with 0.3 °C/min was the best method for
cryopreservation of in vitro derived bovine embryos.
References
Тодорова, Т. (2007). Ин витро матуриране, оплождане и култивиране на говежди ооцити, влияние на изкуствени методи за активация на ооцитите. Дисертация.
Тодорова, Т. (2008). Витрификация на in vitro получени ембриони от говеда. Животновъдни науки, XLV, 6, 52-56.
Arav, A., Yavin, S., Zeron, Y., Natan, D., Dekel, I., & Gacitua, H. (2002). New trends in gamete’s cryopreservation. Molecular and cellular endocrinology, 187(1-2), 77-81.
Dobrinsky, J. R. (2001). Cryopreservation of pig embryos: adaptation of vitrification technology for embryo transfer. Reproduction (Cambridge, England) Supplement, 58, 325-333.
Dobrinsky, J. R. (2002). Advancements in cryopreservation of domestic animal embryos. Theriogenology, 57(1), 285-302.
De Loos, F., Van Vliet, C., van Maurik, P. V., & Kruip, T. A. (1989). Morphology of immature bovine oocytes. Molecular Reproduction and Development, 24(2), 197-204.
Gilchrist, R. B., Zeng, H. T., Wang, X., Richani, D., Smitz, J., & Thompson, J. G. (2015). Reevaluation and evolution of the simulated physiological oocyte maturation system. Theriogenology, 84(4), 656-657.
Greve, T., H. Callesen, Hyttel, P., Avery, B. (1994). From oocyte to calf: in vivo and in vitro. Animal Production and Biotechnology. G. F.Greppiu.G. Enne. Paris, Elsevier Biofutur: 71-91.
Gutierrez, A., Garde, J., Artiga, C. G., Munoz, I., & Pintado, B. (1993). In vitro survival of murine morulae after quick freezing in the presence of chemically defined macromolecules and different cryoprotectants. Theriogenology, 39(5), 1111-1120.
Karlsson, J. O., Eroglu, A., Toth, T. L., Cravalho, E. G., & Toner, M. (1996). Fertilization and development of mouse oocytes cryopreserved using a theoretically optimized protocol. Human Reproduction, 11(6), 1296-1305.
Koruji, S. M., Movahedin, M., & Rezazadeh Valojerdi, M. (2004). Assessment of Epidermal Growth Factor (EGF) Effects on Development of Vitrified Mouse Morulae to the Blastocyst Stage. Iranian Biomedical Journal, 8(2), 77-82.
Kuleshova, L. L., Shaw, J. M., & Trounson, A. O. (2001). Studies on replacing most of the penetrating cryoprotectant by polymers for embryo cryopreservation. Cryobiology, 43(1), 21-31.
Kuleshova, L. L., & Lopata, A. (2002). Vitrification can be more favorable than slow cooling. Fertility and sterility, 78(3), 449-454.
Lane, M., Lyons, E. A., & Bavister, B. D. (2000). Cryopreservation reduces the ability of hamster 2-cell embryos to regulate intracellular pH. Human Reproduction, 15(2), 389-394.
Mazur, P. (1990). Equilibrium, quasi-equilibrium, and nonequilibrium freezing of mammalian embryos. Cell biophysics, 17(1), 53-92.
Massip, A. (2001). Cryopreservation of embryos of farm animals. Reproduction in Domestic Animals, 36(2), 49-55.
Pugh, P. A., Tervit, H. R., & Niemann, H. (2000). Effects of vitrification medium composition on the survival of bovine in vitro produced embryos, following in strawdilution, in vitro and in vivo following transfer. Animal reproduction science, 58(1-2), 9-22.
Rall, W. F., & Fahy, G. M. (1985). Ice-free cryopreservation of mouse embryos at− 196 C by vitrification. Nature, 313(6003), 573.
Seidel, G. E., Elsden, R. P., & Brink, Z. (1990). Cryopreservation of bovine embryos in media with chemically defined macromolecules. Theriogenology, 33(1), 322.
Seidel, G. E., & Walker, D. J. (2006). Pregnancy rates with embryos vitrified in 0.25-mL straws. J. Reprod. Dev, 52, S71-S76.
Vajta, G., Booth, P. J., Holm, P., Greve, T., & Calleson, H. (1997). Successful vitrification of early stage bovine in vitro produced embryos with the open pulled straw (OPS) method.
Wrenzycki, C., Herrmann, D., & Niemann, H. (2007). Messenger RNA in oocytes and embryos in relation to embryo viability. Theriogenology, 68, S77-S83.
Whittingham, D.G., Leibo, S.P., Mazur, P. (1972) Survival of mouse embryos frozen to -196 °C and -269 °C. Science;178:411-414.
Yavin, S., Aroyo, A., Roth, Z., & Arav, A. (2009). Embryo cryopreservation in the presence of low concentration of vitrification solution with sealed pulled straws in liquid nitrogen slush. Human Reproduction, 24(4), 797-804.
Downloads
Published
Issue
Section
Categories
License
Copyright (c) 2018 Bulgarian Journal of Animal Husbandry
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
