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한국재래닭(오계) 원시생식세포의 완만동결과 급속동결의 비교

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

김동훈|김현|도윤정|한재용|고응규|성환후|변미정|김성우
한국축산학회 (Korean Society of Animal Sciences and Technology)|2013.10|한국축산학회지 (Journal of Animal Science and Technology)|제55권 제5호|pp. 417-725

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참고문헌

키워드

생존율ViabilityEG오계원시생식세포Slow freezing급속동결완만동결Primordial germ cells(PGCs)Korean native chicken(Ogye)
주제분류축산학

초록

동결 닭 PGCs의 생식계열 키메라를 이용한 생체에의 복원을 실용화 하기 위해서는, 닭 PGCs의 동결보존기술의 향상에 의해 동결및 융해 후의 많은 생존세포를 확보 하는 것과, 생식계열 키메라의제작효율을 높이는 것이 반드시 필요하다. 닭 PGCs는 배양 5.5일령의 닭 원시생식선으로부터 채취하고, MACS 방법에 의해서 순수 닭 PGCs를 분리했다. 닭 PGCs의 동결보존실험결과 다음의 4종류의 동결방법을 각각 비교 검토했다. 1. 플라스틱 스트로에 의한 완만동결법 (SF), 동결보호물질은 2M 에틸렌 글리콜 (EG), 2.스트로에 의한 급속동결법 (RF), 8M EG + 7% PVP, 3. 동결용Cryotube에 의한 SF, 2M EG, 4. 튜브에 의한 SF, 10% DMSO. 동결 및 융해 후의 PGCs의 생존율은 각각 76.4%, 70.6%, 80.5%, 78.1%로 관찰되었다.

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 2.0M 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.

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