Effects of Cryoprotectant, Warming Solution and Removal of Lipid on Viability of Porcine Nuclear Transfer Embryos Vitrified by Open Pulled Straw Method
Pei-QingCong, Eun-Sook Song, Eui-Sook Kim, Zhao-Hua Li, Yong-Hua Zhang, Jang-Mi Lee, Young-Joo Yi, Chang-Sik Park
언어
영어(ENG)
URL
https://www.earticle.net/Article/A46737
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초록
영어
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℃. 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.
목차
ABSTRACT INTRIDUSTION MATERIALS AND METHODS Collection and In Vitro Maturation of Porcine Oocytes Preparation of Porcine Fetal Fibroblasts Lipid Removal of Porcine Oocyte Nuclear Transfer of Fetal Fibroblast and In Vitro Culture of Porcine Oocytes Evaluation of Blastocyst Produced In Vitro Experimental Design Statistical Analysis RESULTS Effects of Vitrification Solutions on Porcine NT Blastocysts Effects of Warming Solutions on Porcine NT Blastocysts DISCUSSION REFERENCES
키워드
VitrificationCryoprotectantsWarming SolutionLipid removalPorcine nuclear transfer blastocyst
저자
Pei-QingCong [ Research Center for Transgenic Clone Pigs ]
Eun-Sook Song [ Research Center for Transgenic Clone Pigs ]
Eui-Sook Kim [ Research Center for Transgenic Clone Pigs ]
Zhao-Hua Li [ Research Center for Transgenic Clone Pigs ]
Yong-Hua Zhang [ Research Center for Transgenic Clone Pigs ]
Jang-Mi Lee [ Research Center for Transgenic Clone Pigs ]
Young-Joo Yi [ Research Center for Transgenic Clone Pigs ]
Chang-Sik Park [ Research Center for Transgenic Clone Pigs, Division of Animal Science and Resources, Chungnam National University ]
Corresponding author