Journals / Ankara Üniversitesi Veteriner Fakültesi Dergisi / 2026 / Volume: 66 Issue: 1

Effects of equilibration media and co-culture on vitrification of sheep blastocysts derived in vitro

Pages
—
Publication Date
2018-12-31
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

This study shows that vitrification of sheep embryos using ethylene glycol instead of glycerol as a first equilibration cryoprotectant could give reasonable survival rates and that co-culture of embryos with sheep oviduct epithelial cell has no beneficial effect on vitrification of embryos.   

📌 3 Key Takeaways

🎯
Objective & Hypothesis:The effects of two vitrification protocols on the survival of sheep blastocysts were examined in embryos produced in vitro with sheep oviduct epithelial cells co-culture (CC) or without co-culture (C).
📊
Key Findings & Data:Oocytes collected from slaughtered ewes were matured for 24 h, fertilized with fresh ram semen for 20 h and cultured in SOF medium for up to 9 days in vitro.
💡
Conclusion & Impact:This study shows that vitrification of sheep embryos using ethylene glycol instead of glycerol as a first equilibration cryoprotectant could give reasonable survival rates and that co-culture of embryos with sheep oviduct epithelial cell has no beneficial effect on vitrification of embryos.   
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Co-culture · equilibration medium · in vitro fertilization

Abstract

The effects of two vitrification protocols on the survival of sheep blastocysts were examined in embryos produced in vitro with sheep oviduct epithelial cells co-culture (CC) or without co-culture (C). Oocytes collected from slaughtered ewes were matured for 24 h, fertilized with fresh ram semen for 20 h and cultured in SOF medium for up to 9 days in vitro. For vitrification, blastocyst stage embryos were assigned to two equilibration groups (20% ethylen glycol (EG) or 10% glycerol (G) for the first equilibration), and as the second equilibration they were kept in 20% ethylen glycol plus 10% glycerol for 5 minutes. After 30 sec in vitrification solution (25% ethylen glycol + 25% glycerol), they were immediately immersed into liquid nitrogen. After thawing procedure, embryos were transferred into 0.25 M sucrose for 5 min, washed in Hepes buffered synthetic oviduct fluid (HSOF) and cultured in SOF medium for 24 h. Cleavage rates were 75.2% in C and 74.2% in CC groups, and blastocyst rates were 14.4% in C and 17.1% in CC groups. After in vitro culture of vitrified-thawed blastocysts, survival rates were 62.1, 38.4, 30.2, and 39.3% in C-EG, CC-EG, C-G and CC-G groups, respectively. This study shows that vitrification of sheep embryos using ethylene glycol instead of glycerol as a first equilibration cryoprotectant could give reasonable survival rates and that co-culture of embryos with sheep oviduct epithelial cell has no beneficial effect on vitrification of embryos.   

🔖

Cite This Article

Atalla, H., Evecen, M., Pabuccuoğlu, S., Birler, S. (2018). Effects of equilibration media and co-culture on vitrification of sheep blastocysts derived in vitro. Ankara Üniversitesi Veteriner Fakültesi Dergisi, Volume: 66 Issue: 1. https://ajindex.com/articles/1364253

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (31)

  1. Alçay S, Üstüner B, Nur Z (2016): Effects of low molecular weight cryoprotectants on the post-thaw ram sperm quality and fertilizing ability. Small Ruminant Research, 136, 59-64.
  2. Ali J, Shelton J (1993): Successful vitrification of day-6 sheep embryos. Journal of J Reprod Fert, 99, 65-70. 3. Bavister BD (1998): Role of oviductal secretions in embryonic growth in vivo and in vitro. Theriogenology, 29, 143-154.
  3. Birler S, Pabuccuoğlu S, Alkan S, et al. (2001): Effects of different maturation and culture media on IVF of sheep oocytes. Pakistan Vet J, 100-6,102.
  4. De Paz P, Sanchez A, Fernandez J, et al. (1994): Sheep embryo cryopreservation by vitrification and conventional freezing. Theriogenology, 42, 327-338.
  5. Donnay I, Van Langendonckt A, Auquier P, et al. (1997): Effects of co-culture and embryo number on the in vitro development of bovine embryos. Theriogenology, 47, 1549- 1561.
  6. Gandolfi F, Moor R (1987): Stimulation of early embryonic development in the sheep by co-culture with oviduct epithelial cells. J Reprod Fert, 81,23-28.
  7. Garcia-Garcia R, Gonzalez-Bulnes A, Dominguez V, et al. (2006): Survival of frozen-thawed sheep embryos cryopreserved at cleavage stages. Cryobiology, 52, 108- 113.
  8. Gupta MK, Lee HT (2010): Cryopreservation of oocytes and embryos by vitrification. Korean J Reprod Med, 37, 267-291.
  9. Heidari B, Shirazi A, Naderi M, et al. (2013): Effect of various co-culture systems on embryo development in ovine. Czech J Anim Sci, 58, 443-452.
  10. Jang H, Jung Y, Cheong H, et al. (2008): Effects of cell status of bovine oviduct epithelial cell (BOEC) on the development of bovine IVM/IVF embryos and gene expression in the BOEC used or not used for the embryo culture. Asian-Aust J Anim Sci, 21, 980-987.
  11. Ledda S, Bogliolo L, Leoni G, et al. (1995): Two culture systems showing a biphasic effect on ovine embryo development from the 1-2 cell stage to hatched blastocysts. Reprod Nutr Dev, 35, 629-637.
  12. Manokaran S, Veerapandian C, Balasubramanian S (2012): In vitro fertilization of sheep oocytes matured in two different media. Indian J Small Ruminants, 18, 44-46.
  13. Martinez A, Matkovic M (1998): Cryopreservation of ovine embryos: slow freezing and vitrification. Theriogenology, 49, 1039-1049.
  14. Martínez A, Valcárcel A, Furnus C, et al. (2006): Cryopreservation of in vitro-produced ovine embryos. Small Ruminant Research, 63, 288-296.
  15. Nedambale T, Dinnyes A, Groen W, et al. (2004): Comparison on in vitro fertilized bovine embryos cultured in KSOM or SOF and cryopreserved by slow freezing or vitrification. Theriogenology, 62, 437-449.
  16. Niemann H (1991): Cryopreservation of ova and embryos from livestock: current status and research needs. Theriogenology, 35, 109-124.
  17. Paramio MT, Izquierdo D (2014): Current status of in vitro embryo production in sheep and goats. Reprod Dom Anim, 49, 37- 48.
  18. Pereira R, Marques C (2008): Animal oocyte and embryo cryopreservation. Cell and Tissue Banking, 9, 267-277.
  19. Riha J, Machatkova M, Pavlok A (2002): Viability of fresh and frozen transferred IVP bovine embryos. Czech J Anim Sci, 47, 261-267.
  20. Sakkas D, Trounson A, Kola I (1989): In vivo cleavage rates and viability obtained for early cleavage mouse embryos in co-culture with oviduct cells. Reprod Fer Dev, 1, 127-136.
  21. Schiewe M, Rall W, Stuart L, et al. (1991): Analysis of cryoprotectant, cooling rate and in situ dilution using conventional freezing or vitrification for cryopreserving sheep embryos. Theriogenology, 36, 279-293.
  22. Shirazi A, Motaghi E (2013): The in vitro fertilization of ovine oocytes in the presence of oviductal cells and its effect on the expression of zygote arrest 1 (zar1) and subsequent embryonic development. J Reprod Infert, 14, 8-16
  23. Songsasen N, Buckrell B, Plante C, et al. (1995): In vitro and in vivo survival of cryopreserved sheep embryos. Cryobiology, 32, 78-91.
  24. Tominaga K (2004): Cryopreservation and sexing of in vivo-and in vitro-produced bovine embryos for their practical use. Journal of Reproduction and Development, 50, 29-38.
  25. Vajta G (2000): Vitrification of the oocytes and embryos of domestic animals. Anim Reprod, 60, 357-364.
  26. Van Wagtendonk-de Leeuw A, Mullaart E, De Roos A, et al. (2000): Effects of different reproduction techniques: AI, MOET or IVP, on health and welfare of bovine offspring. Theriogenology, 53, 575-597.
  27. Villamil PR, Lozano D, Oviedo J, et al. (2012): Developmental rates of in vivo and in vitro produced bovine embryos cryopreserved in ethylene glycol based solutions by slow freezing or solid surface vitrification. Anim Reprod, 9, 86-92.
  28. Yadav P, Saini A, Kumar A, et al. (1998): Effect of oviductal cell co-culture on cleavage and development of goat IVF embryos. Anim Reprod, 51, 301-306.
  29. Yang N, Lu K, Gordon I, et al. (1992): Vitrification of bovine blastocysts produced in vitro. Theriogenology, 37, 326 (abstract).
  30. Youngs CR (2011): Cryopreservation of preimplantation embryos of cattle, sheep, and goats. J Vis Exp, 54, 2764
  31. Birler S, Pabuccuoğlu S, Alkan S, et al. (2001): Effects of different maturation and culture media on IVF of sheep oocytes. Pakistan Vet J, 100-6,102.