Developmental Competence of Early Stage Porcine Embryos Cultured in Medium with Different Energy Substrate in vitro

Ni Wayan Kurniani Karja(1*), Kazuhiro Kikuchi(2), Mokhamad Fahrudin(3)

(1) Department of Reproduction and Obstetric, Faculty of Veterinary Medicine, Gadjah Mada University,Yogyakarta 55281, Indonesia.Department of Animal Sciences, Reproductive Biology Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan
(2) Department of Animal Sciences, Reproductive Biology Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan
(3) Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor 16680, Indonesia.Department of Animal Sciences, Reproductive Biology Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan
(*) Corresponding Author


To elucidate the effect of energy requirement during the early embryonic development on their developmental ability to develop to blastocyst stage, in vitro fertilized (IVF) porcine one-cell embryos were cultured in modified North Carolina State University (NCSU)-37 supplemented with different energy substrate. Result indicated that the cleavage rate of embryos in Pyr-Lac and Gluc-Pyr-Lact groups was significantly higher than in those in Gluc group and Gluc-Rib group (P < 0.05). At Day 6 of culture, the highest proportion of embryos develop to the blastocyst stage was obtained in the presence of pyruvate-lactate only. In the medium with glucose, the addition of pyruvate-lactate or ribose slightly increased the proportion of embryos develop to the blastocyst stage, however the value were not significantly different form those obtained in the presence of glucose only. The mean cell number in blastocysts derived from Pyr-Lac and Gluc-Pyr-Lact groups were significantly higher than those in the Gluc group (P < 0.05). These results indicated that the presence of glucose only, as energy substrate, during the first 2 days of in vitro culture (IVC) caused a decrease in development of in vitro produced (IVP) porcine embryos to the blastocyst stage and mean cell number in blastocysts.


porcine embryos-energy substrate-in vitro culture

Full Text:



Babior, B.M., 2000. Phagocytes and oxidative stress. Am. J. Med., 109:33-44.

Bavister, B.D., 1979. Fertilization of hamster eggs in vitro at sperm:egg ratios close to unity. J. Exp. Zool., 210:259-264.

Butcher, L., Coates A., Martin, K.L., Rutherford, A.J. and Leese, H.J., 1998. Metabolism of pyruvate by the early human embryo. Biol. Reprod., 58:1054- 1056.

Chatot, C.L., Ziomek, C.A., Bavister, B.D., Lewis, J.L. and Torres, I., 1989. An improved culture medium supports development of random-bred 1-cell mouse embryos in vitro. J. Reprod. Fertil. 86:679-688.

Flood, M.R. and Wiebold, J.L., 1988. Glucose metabolism by preimplantation pig embryos. J. Reprod. Fertil. 84:7-12.

Gandhi, A.P., Lane, M., Gardner, D.K. and Krisher, R. L., 2001 . Substrate utilization in porcine embryos cultured in NCSU23 and G1.2/G2.2 sequential culture media. Mol Reprod Dev. 58:269-275.

Gardner, D.K., Lane, M., and Batt, P., 1993. Uptake and metabolism of pyruvate and glucose by individual sheep preattachment embryos developed in vivo. Mol. Reprod. Dev., 36:313-319.

Guerin, P., El Mouatassim, S. and Menezo, Y., 2001. Oxidative stress and protection against reactive oxygen species in the pre-implantation embryo and its surroundings. Hum. Reprod. Update., 7:175-189.

Houghton, F.D., Thompson, J.G., Kennedy, C.J. and Leese, H.J., 1996. Oxygen consumption and energy metabolism of the early mouse embryo. Mol. Reprod. Dev., 44:476-485.

Johnson, M.H., and Nasr-Esfahani, M.H., 1994. Radical solutions and cultural problems: could free oxygen radicals be responsible for the impaired development of preimplantation mammalian embryos in vitro? Bioessays. 16:31-38.

Karja, N.W.K., Medvedev, S., Onishi, A., Fuchimoto, D., Iwamoto, M., Otoi, T. and Nagai, T., 2004a. Effect of replacement of pyruvate/lactate in culture medium with glucose on preimplantation development of porcine embryos in vitro. J. Reprod. Dev., 50:587-592.

Karja, N.W. K., Wongsrikeao, P., Murakami, M., Agung, B., Fahrudin, M., Nagai, T. and Otoi, T., 2004b. Effects of oxygen tension on the development and quality of porcine in vitro fertilized embryos. Theriogenology. 62:1585-1595.

Kikuchi, K., Onishi, A., Kashiwazaki, N., Iwamoto, M., Noguchi, J., Kaneko, H., Akita, T. and Nagai, T., 2002. Successful piglet production after transfer of blastocysts produced by a modified in vitro system. Biol. Reprod., 66:1033-1041.

Kim, H.S., Lee, G.S., Hyun, S.H., Lee, S.H., Nam, D.H., Jeong, Y.W., Kim, S., Kang, S.K., Lee, B.C. and Hwang, W.S., 2004. Improved in vitro development of porcine embryos with different energy substrates and serum. Theriogenology. 61:1381-1393.

Leese, H.J., 1991. Metabolism of the preimplantation mammalian embryo. Oxf Rev. Reprod. Biol. 13:35-72.

M a n e s , C . , a n d L a i , N . C . , 1 9 9 5 . Nonmitochondrial oxygen utilization by rabbit blastocysts and surface production of superoxide radicals. J. Reprod. Fertil. 104: 69-75.

Nichol, R., Hunter, R.H., Gardner, D.K., Leese, H.J., and Cooke. G.M., 1992. Concentrations of energy substrates in oviductal fluid and blood plasma of pigs during the peri-ovulatory period. J. Reprod. Fertil., 96:699-707.

Petters, R.M., and Wells, K.D., 1993. Culture of pig embryos. J. Reprod. Fertil. Suppl. 48:61-73.

Rieger, D., 1992. Relationship between energy metabolism and development of early mammalian embryos. Theriogenology. 37:75-93.

Salvemini, F., Franze, A., Iervolino, A., Filosa, S., Salzano, S., and Ursini. M.V.,1999. Enhanced glutathione levels and oxidoresistance mediated by increased glucose-6-phosphate dehydrogenase expression. J. Biol. Chem., 274:2750-2757.

Sturmey, R.G., and Leese, H.J., 2003. Energy metabolism in pig oocytes and early embryos. Reproduction. 126:197-204.

Swain, J.E., Bormann, C.L., Clark, S.G., Walters, E.M., Wheeler, M.B., Krisher, R.L., 2002. Use of energy substrates by various stage preimplantation pig embryos produced in vivo and in vitro. Reproduction. 123:253-260.

Telford, N.A., Watson, A.J. and Schultz, G.A., 1990. Transition from maternal to embryonic control in early mammalian development: a comparison of several species. Mol. Reprod. Dev., 26:90-100.

Thompson, J.G.E., Simpson, A.C., Pugh, P. A., and Tervit, H. R., 1992 . Requirements for glucose during in vitro culture of sheep preimplantation embryos. Mol. Reprod. Dev., 31:253- 257.

Thompson, J.G., Bell, A.C., Pugh, P.A. and Tervit, H.R., 1993. Metabolism of pyruvate by pre-elongation sheep embryos and effect of pyruvate and lactate concentrations during culture in vitro. Reprod. Fertil. Dev., 5:417-23.


Article Metrics

Abstract views : 1205 | views : 1431


  • There are currently no refbacks.

Copyright (c) 2015 Indonesian Journal of Biotechnology