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번식생리 및 내분비 : 한국재래닭(오계) 원시생식세포의 완만동결과 급속동결의 비교
김성우 ( Sung Woo Kim ),고응규 ( Yeoung Gyu Ko ),변미정 ( Mi Jeong Byun ),도윤정 ( Yoon Jung Do ),한재용 ( Jae Yong Han ),김동훈 ( Dong Hun Kim ),성환후 ( Hwan Hoo Seong ),김현 ( Hyun Kim ) 한국동물자원과학회(구 한국축산학회) 2013 한국축산학회지 Vol.55 No.5
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 2.0M 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℃ 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.
번식생리 및 내분비 : 한국재래닭(오계) 원시생식세포에 있어 동결방법의 개선이 융해 후생존율에 미치는 영향
김현 ( Hyun Kim ),김동훈 ( Dong Hun Kim ),한재용 ( Jae Yong Han ),최성복 ( Sung Bok Choi ),고응규 ( Yeoung Gyu Ko ),도윤정 ( Yoon Jung Do ),성환후 ( Hwan Hoo Seong ),김성우 ( Sung Woo Kim ) 한국동물자원과학회(구 한국축산학회) 2013 한국축산학회지 Vol.55 No.5
This study was conducted to establish methods for preserving chicken primordial germ cells(PGCs) for long-term storage in liquid nitrogen and for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of fetal bovine serum(FBS) or chicken serum(CS) treatment on the viability of cryopreserved PGCs from Korean Native Chicken (Ogye). PGCs separated from a germinal gonad of an early embryo at day 5.5-6(stage 28) were suspended in a freezing medium containing freezing and protective agents(dimethyl sulfoxide(DMSO), ethylene glycol(EG) and glycerol). The values from 0, 5, 10, and 15 % DMSO plus FBS treatment were 21.6, 30.36, 36.42, 50.39, and 48.36 %, respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% EG+FCS treatment(p<0.05) (64.36% vs. 50.66%). This study establishes a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGC in liquid nitrogen at a germplasm repository and an ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted to improve the production of germline chimeras.