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Effects of Oviductal Fluid, Culture Media and Zona Pellucida Removal on the Development of Porcine Embryos by Nuclear Transfer

  • Zhang, Y.H. (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Song, E.S. (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Kim, E.S. (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Cong, P.Q. (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Lee, S.H. (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Lee, J.W. (Department of Animal Resource Science, College of Industrial Science, Kongju National University) ;
  • Yi, Y.J. (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Park, Chang-Sik (Research Center for Transgenic Cloned Pigs, Chungnam National University)
  • Received : 2008.12.16
  • Accepted : 2009.03.19
  • Published : 2009.07.01

Abstract

The aim of this study was to compare the effects of oviductal fluid, porcine zygote medium (PZM)-3, PZM-4 and PZM-5, and modified PZM-5 culture media, and determine the effects of zona pellucida (ZP) removal on the development of nuclear transfer (NT) embryos. There were no significant differences in the rates of fusion and cleavage among the five different oviductal fluid concentrations. However, the rates of blastocyst formation and the cell numbers per blastocyst were high in the embryos at the 14 and 28 $\mu{g}$/ml concentrations of oviductal fluid compared to the 0, 56 and 100 $\mu{g}$/ml concentrations. The rates of cleavage and blastocyst formation, and the cell numbers per blastocyst were higher in the PZM-3, PZM-5 and modified PZM-5 media than in the PZM-4 medium. However, there were no significant differences in the fusion rates of oocytes among the four culture media. The cell numbers per blastocyst in the embryos without ZP were significantly greater than those with ZP. However, there were no significant differences in the rates of fusion, cleavage and blastocyst formation between the embryos with and without ZP. In conclusion, we improved blastocyst development and the quality of NT embryos by replacing PVA with 3 mg/ml of BSA in PZM-5 medium and supplementing the PZM-5 medium with 14 $\mu{g}$/ml oviductal fluid. The NT embryos produced by the zona-free NT method had a high rate of blastocyst formation in the modified PZM-5 medium.

Keywords

Oviductal Fluid;Nuclear Transfer;Culture Media;Zona Pellucida;Porcine

Acknowledgement

Supported by : Korea Science and Engineering Foundation (KOSEF)

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