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Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos

Year 2016, Volume: 42 Issue: 2, 150 - 155, 04.07.2016
https://doi.org/10.16988/iuvfd.2016.74824

Abstract

In the first stage of the study, oocytes obtained from ovaries from abbatoir (n=2990) were incubated in maturation medium for 24 h. The maturated oocytes were left for in vitro fertilization (IVF) for 20 hours. The cleavaged embryos (n=1305) were left for in vitro culture (IVC) for six days. Then morula-blastocyst stage embryos divided randomly in three groups (Grup I: 0.5°C/min, Grup II: 0.8°C/min, Grup III: 1°C/min). Embryos in each group (n=50) were frozen at different cooling rates in 1.5 M ethylene glycol containing freezing medium. As a result, 0.5°C/min cooling rate group was found as the most successful group (P<0.05). In the second stage of the study, in vivo embryos (morula-blastocyst) derived from donor sheep were frozen at 0.5 °C/min. Frozen-thawed nineteen embryos were transferred to hormonally pretreated seventeen recipients. At the 60th day of the transfer, three recipients were diagnosed as pregnant and one of them had twins. One of the three recipients gave birth and two sheeps had early embryonic loss are observed in the later ultrasound scanning. In the study, 0.5°C/min cooling rate in the slow freezing of sheep embryos was found out to be more successful cooling rates and pregnancy following a healthy lambing is achieved

References

  • Akal, E., Selçuk M., 2013. Spermatozoon DNA’sının hasarına neden olan faktörler, tamir ve hasar tespit mekanizmaları. Journal of the Faculty of Veterinary Medicine Istanbul University 39, 284-293.
  • Atalla, H., 2002. Freezing of sheep embryos produced in vitro. PhD Thesis. Istanbul University, Istanbul, TURKEY,
  • Ataman, M.B., 1998. Koyun embriyosunun dondurulması. Hayvancılık Araştırma Dergisi 8, 38-39.
  • Bari, F., Khalid, M., Haresign, W., Murray, A., Merrell, B., 2003. Factors affecting the survival of sheep embryos after transfer within a MOET program. Theriogenology 59, 1265-1275.
  • Birler, S., Pabuccuoğlu, S., Demir, K, Cirit, U., Karaman, E., Bacınoğlu, S., Evecen, M., Alkan, S., Baran, A., Bakırer, G., Özcan, C., Koçak, Ö., Kılıçarslan, R., Kaşıkçı, G., Toydemir, S., Dinç, H., Koban, E., Togan, İ., İleri, İ.K., Ak, K., 2010. Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients Journal of the Faculty of Veterinary Medicine Istanbul University 36, 1-8.
  • Boland, M.P., Lonergan, P, O’Callaghanz, D., 2001. Effect of nutrition on endocrine parameters, ovarian physiology, and oocyte and embryo development. Theriogenology 55, 1323-1340.
  • Dattena, M., Ptak, G., Loi, P., Cappai, P., 2000. Survival and viability of vitrified in vitro and in vivo produced ovine blastocysts. Theriogenology 53, 1511-1519. Dattena, M., Accardo, C., Pilichi, S., Isachenko, V., Mara, L., Chessa, B., Cappai, P., 2004. Comparison of different vitrification protocols on viability after of ovine blastocysts in vitro produced and in vivo derived. Theriogenology 62, 481-493.
  • Emsen, E., Koşum, N., 2009. Koyunculukta yeni üretim teknikleri. Uludağ Üniversitesi Ziraat Fakültesi Dergisi 23, 33-42.
  • Fowler, A., Toner, M., 2005. Cryo-Injury and Biopreservation. Annals New York Academy of Sciences 1066, 119-135.
  • Garcia-Garcia, R.M, Gonzalez-Bulnes, A., Dominguez, V., Veiga-Lopez, A., Cocero, M.J., 2006. Survival of frozenthawed sheep embryos cryopreserved at cleavage stages. Cryobiology 52, 108-113.
  • Gasparrini, B., 2002. In vitro embryo production in buffalo species. State of art. Theriogenology 57, 237-256.
  • Grazul-Bilska, A.T., Pant, D., Luther, J.S., Borowicz, P.P., Navanukraw, C., Caton, J.S., Ward, M.A., Redmer, D.A., Reynolds, L.P., 2006. Pregnancy rates and gravid uterine parameters in single, twin and triplet pregnancies in naturally bred ewes and ewes after transfer of in vitro produced embryos. Animal Reproduction Science 92, 268-283.
  • Hasler, J.F., Mccauley, A.D., Latrop, W., Foote, R.H., Elizabethtown, P.A., 1987. Effect of Donor-embryorecipient interactions on pregnancy rate in large scale bovine embryo transfer program. Theriogenology 27, 139-168.
  • Isachenko, V., Alabart, L.J., Dattena, M., Naroth, F., Cappai, P., Isachenko, E., Cocero, J.M., Olivera, J., Roche, A., Accardo, C., Krivokharchenko, A., Folch, J., 2003. New technology for vitrification and field (microscope-free) warming and transfer of small ruminant embryos. Theriogenology 59, 1209-1218.
  • Kaidi, S., Van Langendonckt, A., Massip, A., Dessy, F., Donnay, I., 1999. Cellular alteration after dilution of cryoprotective solutions used for the vitrification of in vitro-produced bovine embryos. Theriogenology 52, 515-525.
  • Martinez, A.G., Brogliatti., G.M., Valcarcel, A., de las Heras, M.A., 2002. Pregnancy rates after transfer of bovine embryos: a field trial. Theriogenology 58, 963-972.
  • O’Brien, J.K., Catt, S.L., Ireland, K.A., Maxwell, W.M.C., Evans, G., 1997. In vitro and in vivo developmental capacity of oocytes from prepubertal and adult sheep. Theriogenology 47, 1433-1443.
  • Robinson, J.J., Ashworth, C.J., Rooke, J.A., Mitchell, L.M., McEvoy, T.G., 2006. Nutrition and fertility in ruminant livestock. Animal Feed Science and Technology 126, 259-276.
  • Sağırkaya, H., Bağış, H., 2003. Memeli embriyolarının kriyoprezervasyonu. Uludağ Üniversitesi Veteriner Fakültesi Dergisi 22, 127-135.
  • Sungur, H., Yurdaydın, N., 1991. Sığır embriyosunun dondurulması. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 31, 1-2.
  • Thibier, M., Guerin, B., 2000. Embryo transfer in small ruminants: The method of choice for health control in germplasm exchanges. Livestock Production Science 62, 253-270.
  • Tominaga, K., Iwaki, F., Hochi, S., 2007. Conventional freezing of in vitro-produced and biopsied bovine blastocysts in the presence of a low concentration of Glycerol and sucrose. Journal of Reproduction and Development 53, 443-447.
  • Vajta, G., Holm, P., Kuwayama, M., Booth, P.J., Jacobsen, H., Greve, T., Callesen, H., 1998. Open pulled straw (OPS) vitrification: a new way to reduced cryoinjuries of bovine ova and embryos. Molecular Reproduction and Development 51, 53-58.
  • Vajta, G., Kuwayama, M., 2006. Improving cryopreservation systems. Theriogenology 65, 236-244.
  • Vieira, A.D., Forell, F., Feltrin, C., Rodrigues, J.L., 2007. Instraw cryoprotectant dilution of IVP bovine blastocysts vitrified in hand-pulled glass micropipettes. Animal Reproduction Sience 99, 377-383.
  • Wani, A.R., Khan, M.Z., Sofi, K.A., Lone, F.A., Malik, A.A., Bhat, F.A., 2012. Effect of cysteamine and epidermal growth factor (EGF) supplementation in maturation medium on in vitro maturation, fertilization and culturing of embryos in sheep. Small Ruminant Research 106, 160-164.

Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos

Year 2016, Volume: 42 Issue: 2, 150 - 155, 04.07.2016
https://doi.org/10.16988/iuvfd.2016.74824

Abstract

Çalışmanın ilk bölümünde, mezbahadan sağlanan ovaryumlardan kazanılan oositler (n=2990) olgunlaştırma medyumu içerisinde 24 saat süreyle olgunlaştırıldı. Ardından, 20 saat süreyle İn Vitro Fertilizasyona (İVF) bırakıldılar. Yarıklanma gösteren embriyolar (n=1305), Sentetik Ovidukt Fluid (SOF) medyumu içerisine alınarak altı gün süresince İn Vitro Kültüre (İVK) bırakıldılar. İVK sonrası elde edilen morula-blastosist aşamasındaki embriyolar rastlantısal şekilde üç farklı dondurma hızı grubuna eşit olarak ayrıldılar (Grup I: 0,5 °C /dk, Grup II: 0,8 °C /dk, Grup III: 1 °C /dk). Her bir gruptaki embriyolar (n=50), 1,5 M etilen glikol bulunan dondurma medyumu içerisinde farklı soğutma hızlarında donduruldu. Sonuçta 0,5 °C/dk soğutma hızının en başarılı grup olduğu belirlendi (P<0,05). Çalışmanın ikinci bölümünde, verici koyunlardan elde edilen in vivo embriyolar (Morula-Blastosist) çalışmanın birinci bölümünde bulunan en başarılı soğutma hızı (0,5 °C/dk) ile soğutularak donduruldu. Dondurulan 19 adet embriyo hormonel olarak hazırlanmış 17 alıcı koyuna transfer edildi. Transfer sonrası 60. günde yapılan ultrason muayenesinde üç adet koyunda gebeliklere ait embriyonik keseler gözlendi ve bu koyunlardan birinde de ikiz gebelik saptandı. Gebe koyunlardan bir tanesinde doğum gerçekleşti; diğer iki koyunda ise ileriki dönemde yapılan ultrason muayenesinde gebeliklerin sonlandığı gözlendi. Çalışma sonucunda, koyun embriyolarının dondurulması sırasında 0,5°C/dk soğutma hızının en başarılı hız olduğu ve bu yöntemle dondurulan embriyoların transferinden de gebelik elde edilebildiği saptandı.

References

  • Akal, E., Selçuk M., 2013. Spermatozoon DNA’sının hasarına neden olan faktörler, tamir ve hasar tespit mekanizmaları. Journal of the Faculty of Veterinary Medicine Istanbul University 39, 284-293.
  • Atalla, H., 2002. Freezing of sheep embryos produced in vitro. PhD Thesis. Istanbul University, Istanbul, TURKEY,
  • Ataman, M.B., 1998. Koyun embriyosunun dondurulması. Hayvancılık Araştırma Dergisi 8, 38-39.
  • Bari, F., Khalid, M., Haresign, W., Murray, A., Merrell, B., 2003. Factors affecting the survival of sheep embryos after transfer within a MOET program. Theriogenology 59, 1265-1275.
  • Birler, S., Pabuccuoğlu, S., Demir, K, Cirit, U., Karaman, E., Bacınoğlu, S., Evecen, M., Alkan, S., Baran, A., Bakırer, G., Özcan, C., Koçak, Ö., Kılıçarslan, R., Kaşıkçı, G., Toydemir, S., Dinç, H., Koban, E., Togan, İ., İleri, İ.K., Ak, K., 2010. Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients Journal of the Faculty of Veterinary Medicine Istanbul University 36, 1-8.
  • Boland, M.P., Lonergan, P, O’Callaghanz, D., 2001. Effect of nutrition on endocrine parameters, ovarian physiology, and oocyte and embryo development. Theriogenology 55, 1323-1340.
  • Dattena, M., Ptak, G., Loi, P., Cappai, P., 2000. Survival and viability of vitrified in vitro and in vivo produced ovine blastocysts. Theriogenology 53, 1511-1519. Dattena, M., Accardo, C., Pilichi, S., Isachenko, V., Mara, L., Chessa, B., Cappai, P., 2004. Comparison of different vitrification protocols on viability after of ovine blastocysts in vitro produced and in vivo derived. Theriogenology 62, 481-493.
  • Emsen, E., Koşum, N., 2009. Koyunculukta yeni üretim teknikleri. Uludağ Üniversitesi Ziraat Fakültesi Dergisi 23, 33-42.
  • Fowler, A., Toner, M., 2005. Cryo-Injury and Biopreservation. Annals New York Academy of Sciences 1066, 119-135.
  • Garcia-Garcia, R.M, Gonzalez-Bulnes, A., Dominguez, V., Veiga-Lopez, A., Cocero, M.J., 2006. Survival of frozenthawed sheep embryos cryopreserved at cleavage stages. Cryobiology 52, 108-113.
  • Gasparrini, B., 2002. In vitro embryo production in buffalo species. State of art. Theriogenology 57, 237-256.
  • Grazul-Bilska, A.T., Pant, D., Luther, J.S., Borowicz, P.P., Navanukraw, C., Caton, J.S., Ward, M.A., Redmer, D.A., Reynolds, L.P., 2006. Pregnancy rates and gravid uterine parameters in single, twin and triplet pregnancies in naturally bred ewes and ewes after transfer of in vitro produced embryos. Animal Reproduction Science 92, 268-283.
  • Hasler, J.F., Mccauley, A.D., Latrop, W., Foote, R.H., Elizabethtown, P.A., 1987. Effect of Donor-embryorecipient interactions on pregnancy rate in large scale bovine embryo transfer program. Theriogenology 27, 139-168.
  • Isachenko, V., Alabart, L.J., Dattena, M., Naroth, F., Cappai, P., Isachenko, E., Cocero, J.M., Olivera, J., Roche, A., Accardo, C., Krivokharchenko, A., Folch, J., 2003. New technology for vitrification and field (microscope-free) warming and transfer of small ruminant embryos. Theriogenology 59, 1209-1218.
  • Kaidi, S., Van Langendonckt, A., Massip, A., Dessy, F., Donnay, I., 1999. Cellular alteration after dilution of cryoprotective solutions used for the vitrification of in vitro-produced bovine embryos. Theriogenology 52, 515-525.
  • Martinez, A.G., Brogliatti., G.M., Valcarcel, A., de las Heras, M.A., 2002. Pregnancy rates after transfer of bovine embryos: a field trial. Theriogenology 58, 963-972.
  • O’Brien, J.K., Catt, S.L., Ireland, K.A., Maxwell, W.M.C., Evans, G., 1997. In vitro and in vivo developmental capacity of oocytes from prepubertal and adult sheep. Theriogenology 47, 1433-1443.
  • Robinson, J.J., Ashworth, C.J., Rooke, J.A., Mitchell, L.M., McEvoy, T.G., 2006. Nutrition and fertility in ruminant livestock. Animal Feed Science and Technology 126, 259-276.
  • Sağırkaya, H., Bağış, H., 2003. Memeli embriyolarının kriyoprezervasyonu. Uludağ Üniversitesi Veteriner Fakültesi Dergisi 22, 127-135.
  • Sungur, H., Yurdaydın, N., 1991. Sığır embriyosunun dondurulması. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 31, 1-2.
  • Thibier, M., Guerin, B., 2000. Embryo transfer in small ruminants: The method of choice for health control in germplasm exchanges. Livestock Production Science 62, 253-270.
  • Tominaga, K., Iwaki, F., Hochi, S., 2007. Conventional freezing of in vitro-produced and biopsied bovine blastocysts in the presence of a low concentration of Glycerol and sucrose. Journal of Reproduction and Development 53, 443-447.
  • Vajta, G., Holm, P., Kuwayama, M., Booth, P.J., Jacobsen, H., Greve, T., Callesen, H., 1998. Open pulled straw (OPS) vitrification: a new way to reduced cryoinjuries of bovine ova and embryos. Molecular Reproduction and Development 51, 53-58.
  • Vajta, G., Kuwayama, M., 2006. Improving cryopreservation systems. Theriogenology 65, 236-244.
  • Vieira, A.D., Forell, F., Feltrin, C., Rodrigues, J.L., 2007. Instraw cryoprotectant dilution of IVP bovine blastocysts vitrified in hand-pulled glass micropipettes. Animal Reproduction Sience 99, 377-383.
  • Wani, A.R., Khan, M.Z., Sofi, K.A., Lone, F.A., Malik, A.A., Bhat, F.A., 2012. Effect of cysteamine and epidermal growth factor (EGF) supplementation in maturation medium on in vitro maturation, fertilization and culturing of embryos in sheep. Small Ruminant Research 106, 160-164.
There are 26 citations in total.

Details

Subjects Health Care Administration
Journal Section Research Article
Authors

Elif Karaman Öztürk This is me

Kamber Demir This is me

Ramazan Arıcı

Sema Birler

Serhat Pabuccuoğlu

Mithat Evecen

Publication Date July 4, 2016
Published in Issue Year 2016 Volume: 42 Issue: 2

Cite

APA Karaman Öztürk, E., Demir, K., Arıcı, R., Birler, S., et al. (2016). Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos. İstanbul Üniversitesi Veteriner Fakültesi Dergisi, 42(2), 150-155. https://doi.org/10.16988/iuvfd.2016.74824
AMA Karaman Öztürk E, Demir K, Arıcı R, Birler S, Pabuccuoğlu S, Evecen M. Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos. iuvfd. June 2016;42(2):150-155. doi:10.16988/iuvfd.2016.74824
Chicago Karaman Öztürk, Elif, Kamber Demir, Ramazan Arıcı, Sema Birler, Serhat Pabuccuoğlu, and Mithat Evecen. “Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos”. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 42, no. 2 (June 2016): 150-55. https://doi.org/10.16988/iuvfd.2016.74824.
EndNote Karaman Öztürk E, Demir K, Arıcı R, Birler S, Pabuccuoğlu S, Evecen M (June 1, 2016) Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 42 2 150–155.
IEEE E. Karaman Öztürk, K. Demir, R. Arıcı, S. Birler, S. Pabuccuoğlu, and M. Evecen, “Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos”, iuvfd, vol. 42, no. 2, pp. 150–155, 2016, doi: 10.16988/iuvfd.2016.74824.
ISNAD Karaman Öztürk, Elif et al. “Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos”. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 42/2 (June 2016), 150-155. https://doi.org/10.16988/iuvfd.2016.74824.
JAMA Karaman Öztürk E, Demir K, Arıcı R, Birler S, Pabuccuoğlu S, Evecen M. Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos. iuvfd. 2016;42:150–155.
MLA Karaman Öztürk, Elif et al. “Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos”. İstanbul Üniversitesi Veteriner Fakültesi Dergisi, vol. 42, no. 2, 2016, pp. 150-5, doi:10.16988/iuvfd.2016.74824.
Vancouver Karaman Öztürk E, Demir K, Arıcı R, Birler S, Pabuccuoğlu S, Evecen M. Effect of Different Cooling Rates on Embryo Survivability and Pregnancy Rates in Freezing Sheep Embryos. iuvfd. 2016;42(2):150-5.