CRYOPRESERVATION OF SHEEP OOCYTES AND EMBRYOS USING LOCAL, MODIFIEID VITRIFICATION TOOL
DOI:
https://doi.org/10.36103/ijas.v50iSpecial.192Keywords:
vitrification, viability, oocytes, embryos, sheep.Abstract
The present study was aimed to cryopreserve mature, immature oocytes and in vitro produced embryos in Iraqi sheep using vitrification technique by local, simple and cost effective vitrification tool. This tool is an innovative straw called vitripeace invented, designed and used for the first time. Immature oocytes were aspirated from ovaries of slaughtered ewes and subjected to in vitro maturation and in vitro fertilization programs.The immature, mature oocytes and embryos were vitrified using Vitripeace tools, then thawed and assessed for the morphology and viability. The results revealed non-significant effect of time on viability (%) and normal morphology (%) of vitrified immature and mature oocytes for post-thawing and 2 hours post-thawing. The results showed significant (P<0.05) reduction in the viability (%) of 2 cell embryo namely 88.89% and 77.78 % for post-thawing and two hours post-thawing respectively. The results revealed a significant (P<0.05) reduction on normal morphology of 1 cell embryo namely 88.24 % and 76.47 % for post-thawing and two hours post-thawing respectively. Significant (P<0.05) differences in the percentage of normal morphology were found at post-thawing period for all stages of embryo development which were 90.74%, 88.31, 88.24 and 83.33 for immature , mature oocytes, 1 cell and 2 cell embryos, respectively while no significant differences in the viability at post-thawing period among all stages of embryo development. It was concluded that, successful vitrification of oocytes and embryos was resulted using Vitripeace which was novel, simple and cost effective vitrification tool.
References
Abd-Allah, S.M. 2010. Effects of storage conditions of dromedary camel ovaries on the morphology, viability and development of antral follicular oocytes. Anim. Reprod., 7:65 69.
Al-Hasani, S ., B.Ozmen and N. Koutlaki. 2007. Three years of routine vitrification of human zygotes: is it still fair to advocate slow-rate freezing. Reprod. BioMed. Online, 14: 288-293
Bromfield, J ., G. Coticchio., K.Hutt., R. Sciajno., A. Borini and D. Albertini. 2009. Meiotic spindle dynamics in human oocytes following slow-cooling cryopreservation. Hum. Reprod ., 24: 2114–2123
Chian, R.C., M. Kuwayama., L. Tan., J. Tan., O. Kato and T. Nagai.2004. High survival rate of bovine oocytes matured in vitro following vitrification. J. Reprod. Dev., 50 (6): 685-96.
De Felici, M. and G. Siracusa. 1982.Spontaneous hardening of the zona pellucida of mouse oocytes during in vitro culture. Gamete. Res ., 6: 107-113.
DeSmedt, V ., N. Crozed and A.M .Ahmed.1992. In vitro maturation and fertilization of goat oocytes. Theriogenlogy. 1992; 37:1049-
Dike, I.P. 2009. Efficiency of intracellular cryoprotectants on the cryopreservation of sheep oocytes by controlled slow freezing and vitrification techniques. Journal of Cell and Animal Biology, 3 (3): 044- 049
Fakhrildin, M.B.M. and R.H. Al Moussawi. 2013. Effect of two types and two concentrations of cryoprotectants on ovine oocytes morphology and viability post-vitrification. Iraqi Journal of Embryos and Infertility Researches, 3(6):32-37
Fujiwara, K ., D. Sano., Y. Seita ., T. Inomata.,J.Ito and Kashiwazaki.2010. Ethylene glycol-supplemented calcium free media improve zona penetration of vitrified rat oocytes by sperm cells. J. Reprod. Dev.,56:169-17.10.Ghorbani, M., R. Sadrkhanlou., V. Nejati, , A. Ahmadi, and G.Tizroo.2012. The effects of dimethyl sulfoxide and ethylene glycol as vitrification protectants on different cleavage stages of mouse embryo quality Veterinary Research Forum, 3 (4) 245-249
Herring, J.E.G.2008. The Development and Analysis of A closed System of Vitrification for Mammalian Embryos. A dissertation .the Graduate School of Clemson University, USA, pp:1
Kuwayama, M ., G. Vajta., S. Ieda and O. Kato.2005a. Comparison of open and closed methods for vitrification of human embryos and the elimination of potential contamination. Reprod. BioMed. Online ., 11(5) 608-614
Nedambale, T.L ., F. Du., J. Xu ., X.C. Tian and X Yang.2006. Effects of vitrification and post-thawing interval on the cytoskeleton and subsequent fertilization rate of in vitro derived bovine oocytes. South African Journal of Animal Science, 36 (5):42 -45
Oktay, K ., A.P. Cil., and H. Bang.2006. Efficiency of oocyte cryopreservation: a meta analysis. Fertil. Steril ., 86: 70-80
Pereira, R.M and C.C. Marques.2008. Animal oocyte and embryo cryopreservation. Cell and Tissue Banking, 9: 267-277
Purohit, G.N ., H.Meena, and K.Solanki.2011.Effects of vitrification on immature and in vitro matured, denuded and cumulus compact goat oocytes and their subsequent fertilization. Journal of Reproduction and Infertility, 13(15):53-59
SAS. 2012. SASSTAT User’s Guide for Personal Computers. Release 9.1 SAS Institute Inc., Cary, N. C., USA
Ubaldi, F ., R. Anniballo., S.Romano., E. Baroni., L. Albricci, and S. Colamaria.2010. Cumulative ongoing pregnancy rate achieved with oocyte vitrification and cleavage stage transfer without embryo selection in a standard infertility program. Hum. Reprod ., 25 (5): 1199-1205
Zhou, X.L ., A. Al- Naib., D.W. Sun and P. Lonergan.2010. Bovine oocyte vitrification using the Cryotop method: effect of cumulus cells and vitrification protocol on survival and subsequent development. Cryobiology, 61 (1): 66- 72
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
2.jpg)
2.jpg)
