Effect of Adding Insulin Transferrin Selenium (ITS) in the Medium on Maturation and Fertilization Rates of Bali Cattle Oocytes

Hikmayani Iskandar, H Sonjaya, M Yusuf


The aim of this study was to determine the effect of adding Insulin Transferrin Selenium (ITS) in the medium on maturation and fertilization rates of Bali cattle oocytes. The ovarium of Bali cows were sliced to produce oocytes, then the collected oocytes were subsequently selected based on their quality. Those oocytes were maturated for 24 hours and then fertilized for 18 hours in the incubators of 5% CO2 with temperature of 38.5ºC. Oocytes were stained using 2% of aceto orcein, and observed under a microscope. This study was divided into four treatments at different addition of ITS in the medium and replicated four times, and analized using a Factorial Complete Randomized Design (CRD)at each treatment. The treatments consisted of P0 as control, P1 (5 ng/ml), P2 (10 ng/ml), and P3 (15 ng/ml). Parameters observed in this study were the stages of oocytes maturation rate that consisted of germinal vesicle (GV), germinal vesicle break down (GVBD), metaphase I (M-I), and metaphase II (M-II), and fertilization rate that consisted of absence of pronucleus (P0), 1 pronucleus (PN1), 2 pronucleus (PN2), and more than two pronucleus (>2PN). The results of this study showed that addition of ITS at 5-15 ng/ml did not effect maturation and fertilization rates of Bali cows oocytes.The conculision of the study is addition of ITS at 5 ng / ml produces the best level of maturation and for the best level of fertilization with addition ITS at 15 ng / ml.


Bali cattle ovary, Insulin Transferrin Selenium, maturation, fertlization


Cetica PD, LN. Pintos LN, GC. Dalvit GC, MT. Beconi MT. 2001. Antioxidant enzyme activity and oxidative stress in bovine oocytes in vitro maturation (Abstact). CNBI Jan; 51 (1): 57-64.

Kim S, Lee GS, Lee SH, Kim HS, Jeong YW, Kim JH, Kang SK, Lee BC, Hwang WS. 2005. Embryotropic effect of insulin-like growth factor (IGF)-I and its receptor on development of porcine preimplantation embryos produced by in vitro fertilization and somatic cell nuclear transfer. Mol Reprod Dev. 72: 88-97.

Lee, MS., SK. Kang, B.C. Lee, W.S. Hwang.2005. The beneficial effects of insulin and metformin on in vitro developmental potential of porcine oocytes and embryos.J. BiolReprod 73: 1264-1268.

Wu, S.H., oldfield, J.E., Whanger, P.D., Weswig, P.H., 1973. Effect of selenium, vitamin E, and antioxidant on testicular function in rats.Biol. Reprod. 8, 625-629.

Gutteridge, J.M., 1986. Antioxidant properties of the proteins caeruloplasmin, albumin and transferring. A study of their activity in serum and synovial fluid from patients with rheumatoid arthritis.Biochim.Biophys. Acta 869, 119-127.

De La Fuente, R., M.J. O’Brien and J.J. Eppig. 1999. Epidermal growth factor enhances preimplantation developmentalbcompetence of maturing mouse oocytes. Hum. Reprod. 14, 3060-8.

Gao, Min-zhi, Yu-bao Wang and Xiao-yun Wu. 2007. In Vitro Maturation of Immature Oocytes from Preantral Follicles in Prepubertas Mice. Journal of reproduction and Contraception. 18 (1): 25-32

Herrick, J.R., E. Behboodi, E. Memili, S. Blash, Y. Echelard and R.L. Krisher. 2004. Effect of macromolecule supplementation during in vitro maturation of goat oocytes on developmental potential. Mol. Reprod.Dev. 69, 338-46.

Jeong, Y.W., M.S. Hossein, D.P. Bhandari, Y.W. Kim, J.H. Kim, S.W. Park, E. Lee, S.M. Park, Y.I. Jeong, J.Y. Lee, S. Kimand W.S. Hwang. 2008. Effects of insulin-transferrin-selenium in defined and porcine follivular fluid upplemented IVM media on porcide IVF and SCNT embryo production. Anim. Reprod. Sci. 106, 13-24.

Djuwita, I., Riyacumala, V., Mohamad, K., Prasetyaningtijas, WE., dan Nurhidayat. 2012. Pertumbuhan dan Sekresi Protein Hasil Kultur Primer Sel-sel Serebrum Anak Tikus. Jurnal Veteriner. 13(2):125-135.

De Loos FD, Vliet CV, Maurik PV, Kruip ThAM. 1989. Morphology of immature bovine oocytes. Gamete Res 24: 197-204.

Loos de F, Vliet CV., Maurik VP and Kruip ThAM. 1989. Morphology of Immature Bovine Oocytes. Gamete Res. 24: 197-204.

Lapathihis G, Miksik I, Pavlok A, and Moltik J. 2002.Improvement of In Vitro Maturation System for Bovine oocytes and Intracelluler Glutathione Content.http://www.iapg.cas.cz/uzfg/u2.htm. (25 Juni 2017).

Boediono A and Suzuki T. 1996.In Vitro Development of Holste in and Japanese Black Breeds Embryo.Media Veteriner 3: 3-9.

Zeng, Y.S. & M.A. Sirard. 1992. The Effect Of Sera, Bovine Serum Albumin and Follicular Cells On In Vitro Maturation And Fertilization of Porcine oocytes. Theriogenology 37: 779- 790.

Moltik J and Fulka J. 1976. Breakdown and Developmental in Pig oocytes In Vivo and In Vitro. Reprod. Fert.

Tsafriri A. 1985. Oocytes Maturation in Mammals. In: RE Jones, editor. The Vertebrata Ovary. New York: Plenum Press, pp: 27-37.

Djuwita I, Boediono A dan Mohammad K. 2000.Bahan Kuliah Embriology. Bogor: Laboratorium Embriologi Bagian Anatomi FKH IPB. Pp: 10: 1-8.

Rusiyantono, Y., I. Djuwita, B. Purwantara dan Y. Sukra. 2000. The influence of ewe serum on in vitro oocytes maturation and early development of ovine embryos. Media Vet. 7: 13-16.

Djuwita, I., B. Purwantara, M. Fahrudin and Y. Sukra. 1995. The effect of superovulated cow serum on in vitro maturation and fertilization in sheep. Symp.Biotechnol.Anim. Reprod. Bogor, 1 Agustus 1995. Faculty of Veterinary Medicine, Bogor Agricultural Institute. Bogor: 20-22.

Orsi NM, Leese HJ. 2004. Amino acid metabolism of preimplantation bovine embryos cultured with bovine serum albumin or polyvinyl alcohol. Theriogenology.61:561-572.

Boediono A, Rusiyantono Y, Mohamad K, Djuwita I, Harliatien. 2000. Developmental competence of caprine oocytes after in vitro maturation, fertilization and culture. Media Veteriner. 7 (4): 11-17.

Crozet, N., M.A. Ali and M.P. Dubos. 1995. Developmental competence of goat oocytes from follicles of different size categories following maturation, fertilization and culture in vitro. J. Reprod. Fertil. 103: 293-298.

Nadir, S., R.G. Saacke, J. Bame, J. Mullins and S. Degelos. 1993. Effect of freezing semen and dosage of sperm on number of accesory sperm, fertility and embryo quality in artificially inseminated cattle. J. Anim. Sci. 71: 199-204.

Long, C.R., P. Damiani, C.P. Correia, R.A. Maclean, R.T. Duby and J.M. Robl. 1994. Morphology and subsequent development in culture of bovine oocytes matured in vitro under various conditions of fertilization. J. Reprod. Fertil. 102: 361-369.

Dandekar, P. and P. Talbot. 1992. Perivitelline space of mammalian oocytes extra-cellular matrix of unfertilized oocytes and formation of a cortical granule envelope following fertilization. Mol. Reprod. Develop. 31: 135- 143.

Kim, B., Sang, C., Kwang, S., Bok, K., Chang-Hee, H., Jong-Heung, K., Chae-Sik, L., 2002, Effect of Medium Milieu on Sperm Penetration and Pronuclear Formation of Bovine oocytes Matured In vitro. Theriogenology. 57: 2093–2104.


  • There are currently no refbacks.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.