In Vitro Bovine Embryo Production: The Role of FCS and Cysteamine on Cleavage Rate

Yıl 2020, Cilt: 7 Sayı: 2, 136 - 139, 30.06.2020

Asiye İzem Sandal

https://doi.org/10.34087/cbusbed.624912

Öz

The aim of this study was to investigate the effects of Cysteamine (C)
and Fetal Calf Serum (FCS) on cleavage rates in bovine embryo production. The oocytes were obtained from the slaughtered ovaries by slicing
method. They were selected and divided into two groups and maturated in TCM and
TCM+Cysteamine for 24 hour at 38.8ºC in atmosphere of 5% CO₂ in
humidified air. The matured oocytes were
transferred to IVF-TALP and allowed to fertilize with frozen bull semen to be 400-600x10^3/well which
were washed with Sperm-TALP in an incubator containing 5% CO₂ for
18-22 hours at 38.8ºC. At the end of this period, the embryos of each group
which were denuded by vortexing transferred to Synthetic Oviduct Fluid (SOF)
and Charles and Rosenkrans (CR1) media to be developed. These culture media
were divided into 4 subgroups according to whether they contain Cysteamine (C)
and Fetal Calf Serum (FCS). For this reason Group A1: (SOF C+F+), Group
A2: (SOF C+F-), Group A3: (SOF C-F+), Group A4: (SOF C-F-), Group B1: (CR1
C+F+), Group B2: (CR1 C+F-), Group B3: (CR1 C-F+), Group B4: (CR1 C-F-) were
formed. Embryos were cultured in an incubator containing 5% CO₂, 5%
N₂ and 90% humidity at 38.8ºC for 48 hours. At the end of this
period, the clevage rate of the embryos were recorded and there was no
statistical difference (P<0.05) between all groups.

Anahtar Kelimeler

bovine, cleavage, cysteamine, embryo, fetalcalfserum

Kaynakça

• Kitagawa, Y, Suzuki, K, Yoneda, A, Watanabe, T. 2004. Effects of oxygen concentration and antioxidants on the in vitro developmental ability, production of reactive oxygen species (ROS), and DNA fragmentation in porcine embryos. Theriogenology; 62: 1186-1197.
• Aitken, RJ, Harkiss, D, Buckingham, D. 1993. Relationship between iron-catalysed lipid peroxidation potential and human sperm function. Journal of Reproduction and Fertility; 98: 257-265.
• Yuan, YQ, Van Soom, A, Coopman, FOJ, Mintiens, K, Boerjan, ML, Van Zeveren, A, De Kruif, A, Peelman, LJ. 2003. İnfluence of oxygen tension on apoptosis and hatching in bovine embryos cultured in vitro. Theriogenology; 59: 1585-1596.
• Thompson, JG., 2000. In vitro culture and embryo metabolism of cattle and sheep embryos: A decade of achievement. Animal Reproduction Science 60-61, 263-275.
• Raty, M, Ketoja, E, Pitkanen, T, Ahola, V, Kananen, K, Peippo, J. 2011. In vitro maturation supplements affect developmental competence of bovine cumulus-oocyte complexes and embryo quality after vitrification. Cryobiology; 63: 245-255.
• Balasubramanian, S, Rho, GJ. 2007. Effect of cysteamine supplementation of in vitro matured bovine oocytes on chilling sensitivity and development of embryos. Animal Reproduction Science; 98: 282-292.
• Hong, J, Lee, E. 2007. Intrafollicular amino acid concentration and the effect of amino acids in a defined maturation medium on porcine oocyte maturation, fertilization, and preimplantation development. Theriogenology; 68: 728-735.
• Kuran, M, Robinson, JJ, Staines, ME, McEvoy, TG. 2001. Development and de novo protein synthetic activity of bovine embryos produced in vitro in different culture systems. Theriogenology; 55: 593-606.
• Gomez, E, Diez, C. 2000. Effects of glucose and protein sources on bovine embryo development in vitro. Animal Reproduction Science; 58: 23-37.
• Rooke, JA, Ewen, M, Mackie, K, Staines, ME, McEvoy, TG, Sinclair, KD. 2004. Effect of ammonium chloride on the growth and metabolism of bovine ovarian granulosa cells and the development of ovine oocytes matured in the presence of bovine granulosa cells previously exposed to ammonium chloride. Animal Reproduction Science; 84: 53-71.
• Lane, M, Gardner, DK. 1994. Increase in postimplantation development of cultured mouse embryos by amino acids and induction of fetal retardation and exencephaly by ammonium ions. Journal of Reproduction and Fertility; 102: 305-312.
• Fukui, Y, Lee, ES, Araki, N. 1996. Effect of medium renewal during culture in two different culture systems on development to blastocysts from in vitro produced early bovine embryos. Journal of Animal Science; 74: 2752-2758.
• Sovernigo, TC, Adona, PR, Monzani, PS, Guemra, S, Barros, FDA, Lopes, FG, Leal, CLV. 2017. Effects of supplementation of medium with different antioxidants during in vitro maturation of bovine oocytes on subsequent embryo production. Reproduction in Domestic Animals; 52; 561-569.
• Sandal, Aİ, Özdaş, ÖB. 2015. Vitrification of in vitro-produced bovine embryos matured in modified TCM-199 medium. Turkish Journal of Veterinary and Animal Sciences; 39: 688-692.
• Enginler, SO, Özdaş, ÖB, Sandal, AI, Arıcı, R, Ertürk, E, Baran, A, Toydemir, TFS, Tek, Ç, Kılıçarslan, MR, Ak, K. 2016. The Effect Of Cysteamine And Oviductal Cells In Different Culture Media On The Development Of Sheep Embryos. Journal Of The Faculty Of Veterinary Medicine, Kafkas University; 22(1): 139-145.
• Sağırkaya, H, Mısırlıoğlu, M, Kayaç, A, First, NL. Parrish, JJ, Memili, E. 2007. Developmental potential of bovine oocytes cultured in different maturation and culture conditions. Animal Reproduction Science; 101: 225-240.
• Hajarian, H, Aghaz, F, Karami Shabankareh, H. 2017. Replacement of serum with sericin in in vitro maturation and culture media: Effects on embryonic developmental competence of Sanjabi sheep embryo during breeding season. Theriogenology; 92: 144-148.
• Do, VH, Walton, S, Taylor-Robinson, AW. 2016. Improvements to in vitro culture media for use in bovine. Journal of Veterinary Science & Animal Husbandry; 4: 205.
• Sudano, MJ, Paschoal, DM, Rascado, T.daS, Magalhães, LCO, Cromoco, LF, Lima-Neto, JF de, Landim-Alvarenga, F da C. 2011. Lipid content and apoptosis of in vitro-produced bovine embryos as determinants of susceptibility to vitrification Theriogenology; 75: 1211-1220.