초자화 동결과 1-단계 융해된 체세포 핵이식란의 직접 이식 기술로 제주흑우 복제소 생산

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 35권 / 1호
ㆍ저자명 : 김은영, 박민지, 김재연, 박효영, 노은지, 노은형, 송동환, 오창언, 김영훈, 문성호, 이동선, 고문석, 류기중, 박세필

목차

ABSTRACT 서론 재료 및 방법 소 난자의 회수 및 체외성숙 제주흑우 체세포 핵이식배아의 생산 및 체외 배양 체세포 핵이식 배반포기배의 초자화 동결 1-단계 융해 간편법(One-step Dilution) 수란우 발정동기화, 1-단계 융해 체세포 핵이식 배반포기배이식 및 복제소 생산 친자 감별 마커를 이용한 복제 유무 검정 통계처리 결과 초자화 동결 체세포 핵이식 제주흑우 배반포기배의 1-단계융해 후 체외발달율 조사 초자화 동결 체세포 핵이식 제주흑우 배반포기배의 1-단계융해 후 직접이식에 따른 임신 결과 친자 감별 유전자 분석에 의한 복제소의 검정 고찰 인용문헌

영어 초록

One-step dilution and direct transfer would be a practical technique for the field application of frozen embryo. This study was to examine whether Jeju Black Cattle (JBC, Korean Cattle) can be successfully cloned from vitrified and one-tep diluted somatic cell nuclear transfer (SCNT) blastocyst after direct transfer. For vitrification, JBC-SCNT blastocysts were serially exposed in glycerol (G) and ethylene glycol (EG) mixtures〔10% (v/v) G for 5 min., 10% G plus 20% EG (v/v) for 5 min., and 25% G plus 25% EG (v/v) for 30 sec.〕which is diluted in 10% FBS added D-PBS. And then SCNT blastocysts were loaded in 0.25 ml mini straw, placed in cold nitrogen vapor for 3 min. and then plunged into LN2. One-step dilution in straw was done in 25℃ water for 1 min, by placing vertically in the state of plugged- end up and down for 0.5 min, respectively. When in vitro developmental capacity of vitrified SCNT blastocyst was examined at 48 h after one-step dilution, hatched rate (56.4%) was slightly lower than that of control group (62.5%). In field trial, when the vitrified-thawed SCNT blastocysts were transferred into uterus of synchronized 5 recipients, a cloned female JBC was delivered by natural birth on day 299 and healthy at present. In addition, when the short tandem repeat marker analysis of the cloned JBC was evaluated, microsatellite loci of 11 numbers was perfectly matched genotype with donor cell (BK94-14). This study suggested that our developed vitrification and one-step dilution technique can be applied effectively on field trial for cloned animal production, which is even no longer in existence.

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