Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Effects of Cryoprotectant, Warming Solution and Removal of Lipid on Viability of Porcine Nuclear Transfer Embryos Vitrified by Open Pulled Straw Method

  • Cong, Pei-Qing (Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Song, Eun-Sook (Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Kim, Eui-Sook (Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Li, Zhao-Hua (Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Zhang, Yong-Hua (Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Lee, Jang-Mi (Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Yi, Young-Joo (Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Park, Chang-Sik (Research Center for Transgenic Cloned Pigs Chungnam National University)
  • Published : 2007.06.30
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Abstract

This study was carried out to investigate the effects of cryoprotectants, warming solution and removal of lipid on open pulled straw vitrification (OPS) method of porcine embryos produced by nuclear transfer (NT) of fetal fibroblasts. All solutions used during vitrification were prepared with holding medium consisting of 25 mM Hepes buffered TCM199 medium containing 20% fetal bovine serum (FBS) at $38.5^{\circ}C$. The blastocysts derived from NT with or without lipid were vitrified in each medium of different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Also, blastocysts after cryopreservation were warmed into different concentrations of sucrose in warming solution. The optimal concentrations of cryoprotectants in vitrification solution were 10% DMSO + 10% EG in vitrification solution 1 (VS1) and 20% DMSO + 20% EG in vitrification solution 2 (VS2). The optimal concentrations of sucrose were 0.3 M sucrose in warming solution 1 (WS1) and 0.15 M sucrose in warming solution 2 (WS2). lipid removal from oocytes before NT enhanced the viability of NT embryos after vitrification. Our results show that use of the OPS method in conjunction with lipid removal provides effective cryopreservation of porcine nuclear transfer embryos.

Keywords

References

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