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Comparison of Vitrification and Slow Freezing for the Cryopreservation of Chicken Primordial Germ Cell (Ogye)

한국재래닭 (오계) 원시생식세포의 완만동결과 급속동결의 비교

  • Kim, Sung Woo (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Ko, Yeoung-Gyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Byun, Mijeong (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Do, Yoon Jung (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Han, Jae Yong (WCU Biomodulation Major, Department of Agricultural Biotechnology, Seoul National University) ;
  • Kim, Dong Hun (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Seong, Hwan-Hoo (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Kim, Hyun (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 변미정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 도윤정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 한재용 (서울대학교 동물자원과학과) ;
  • 김동훈 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 성환후 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김현 (농촌진흥청 국립축산과학원 가축유전자원시험장)
  • Received : 2013.06.17
  • Accepted : 2013.09.13
  • Published : 2013.10.31

Abstract

We sought to provide a method for freezing and preserving primordial germ cells, or an avian germ cell of a bird, as a material for developmental engineering or species preservation. The aim of this study was to compare the efficacy of slow freezing with a vitrification method for the cryopreservation of chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonad of day 5.5-6 day (stage 28) cultured chick embryos, using the MACS method, were classified into two groups: slow freezing and vitrification. We examined the viability of PGCs after Cryopreservation. Four freezing methods were compared with each other, including the following: Method 1: The PGCs were frozen by a programmed freezer in a plastic straw, including 2.0 M ethylene glycol (EG) as cryoprotective additive (slow freezing) Method 2: The PGCs were vitrified in a plastic straw, including 8.0 M EG, plus 7% polyvinylpyrrolidone (PVP) (rapid freezing). Method 3: The slow freezing was induced with a cryotube including 2.0 M EG Method 4: The PGCs were frozen in a cryotube including 10% dimethyl suloxide (DMSO) (rapid freezing). After freezing and thawing, survival rates of the frozen-thawed PGCs from Method 1 to 4were 76.4%, 70.6%, 80.5% and 78.1% (p<0.05), respectively. The slow freezing ($-80^{\circ}C$ programmed freezer) method may provide better survival rates of frozen-thawed PGCs than the vitrification method for the cryopreservation of PGCs. Therefore, these systems may contribute to the cryopreservation of a rare avian species.

Keywords

Primordial germ cells (PGCs);EG;Viability;Korean native chicken (Ogye);Slow freezing

Acknowledgement

Supported by : 농촌진흥청

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