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RISS 인기검색어
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- 저자 : 김수헌(S. H. Kim) (검색결과 6 건)
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H-Y 항체에 의한 토끼배의 성조절에 관한 연구 1 . 배의 발달과 형광 발현에 의한 자 웅 수정란의 분리
이창규(C . K . Lee),정구민(K . M . Chung),김수헌(S . H . Kim),임경순(K . S . Im) 한국축산학회 1990 한국축산학회지 Vol.32 No.7
Antisera to histocompatibility (H-Y) antigen were used to immunologically presume the sex of rabbit embryos. H-Y antisera were prepared in inbred SD female rat by repeated immunization of spleen cells from males of same strain. The titre of H-Y antibody in antiserum was examined by mouse sperm cytotoxicity and biological tests. Experiments applied delaying ability of development of embryos in H-Y antiserum and binding ability of FITC labelled second antibody. After culture, embryos were observed their morphological characteristics under phase contrast microscope and detected fluorescence on embryos under fluorescence microscope. After detection of fluorescence, embryos were transfered to normal medium and observed their morphological characteristics. 1. When rabbit morula were treated with H-Y antiserum only, the rate of developed and delayed embryos was 47.2 and 52.8% respectively, and the rate of non-fluorescing and fluorescing embryos was 51.4 and 48.6%, respectively. 2. When rabbit morula were cultured in H-Y antiserum followed by complement, the rate of non-fluorescing and fluorescing embryos was 53.6 and 46.4%. respectively. 3. After detection of fluorescence, the embryos were cultured in normal medium. When embryos were treated with H-Y antiserum only, the rate of arrested and developed embryos was 20.8 and 79.2% respectively. However, when embryos were treated with H-Y antiserum followed by complement, the rate of arrested and developed embryos was 42.9 and 57.1% respectively.
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비투과성 동해방지제 Trehalose 가 급속동결한 생쥐상실배의 생존성에 미치는 영향
김수헌(S . H . Kim),정구민(K . M . Chung),이창규(C . K . Lee),임경순(K . S . Im) 한국축산학회 1989 한국축산학회지 Vol.31 No.12
This experiment was carried out to investigate effects of trehalose and sucrose concentration as a nonpermeating cryoprotectant, and equilibration time at room temperature on in vitro development of mouse morula frozen by direct plunging from room temperature to liquid nitrogen The results obtained from this experiment are as follows: 1. When the morula were frozen-thawed ultrarapidly in 3.5M glycerol and 0.25 or 0.50M sucrose, the proportions of morphologically normal embryos and embryos developed to expanded blastocysts were slightly better in 0.50M than 0.25M, but, there was no difference in recovery rate of embryo between two treatments. 2. When the morula were frozen-thawed ultrarapidly in 3.5M glycerol and 0.25 or 0.50M trehalose, the proportions of embryos developed to expanded blastocysts were slightly better in 0.50M than 0.25M, but, there was no difference in recovery rate and the proportions of morphologically normal embryos between two treatments. 3. When the morula were frozen-thawed ultrarapidly in 3.5M glycerol and 0.50M sucrose or trehalose, the proportions of embryos developed to expanded blastocysts were slightly better in trehalose than sucrose. 4. In the equilibration time at room temperature, the proportions of embryos developed to expanded blastocysts were higher at 2..5 to 3 minutes than at shorter( (2minutes) or longer(5 to 10 minutes) minutes. 5. When mouse morula were equilibrated for 2.5 to 3 minutes in 3.5M glycerol and 0.50M trehalose at room temperature and frozen by direct plunging into liquid nitrogen, more than 85% embryos developed to expanded blastocysts after culture
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2단계 급속동결 및 초자화 동결한 토끼상실배의 체외생존성에 관한 연구
정구민,이창규,임경순,김수헌 서울대학교농과대학농업개발연구소 1990 서울대농학연구지 Vol.15 No.1
본 시험은 수정란 급속동결보존기술의 개발을 위한 기초자료를 얻기 위하여 2단계 급속동결 및 초자화동결이 토끼 상실배의 체외발생등에 미치는 영향을 검토하고자 실시하였으며, 얻어진 결과는 다음과 같다. 1. 토끼 상실배를 1.5, 2.0, 2.5 및 3.0 M glycerol과 0.5 M sucrose가 포함된 동결액에 실온에서 10분간 노출후 -30℃에서 30∼40분간 정치하여 급속동결하였을 때 발생율은 각각 36.4, 83.3, 92.3 및 84.2%로 2.5 M glycerol에서 가장 높았다. 2. 토끼 상실배를 1.5, 2.0, 2.5 및 3.0 M 1,2-propanediol과 0.5 M sucrose가 포함된 동결액에 실온에서 10분간 노출 후 -30℃에서 30∼40분간 정치하여 급속동결하였을 때 발생율은 각각 26.6, 55.6, 65.0 및 52.9%로 2.5 M 1,2-propanediol에서 가장 높았으나, glycerol을 사용했을 때 보다 낮았다. 3. 토끼 상실배를 2.5, 3.0 및 3.5 M glycerol과 0.5 M trehalose가 포함된 동결액에 실온에서 10분간 노출후 액체질소에 침지하여 초급속동결한 결과 회수율은 각각 87.5, 92.5 및 92.5%, 형태적으로 정상인 수정란의 비율은 각각 37.5, 55.5 및 60.0%, 그리고 발생율은 각각 13.3, 36.4 및 37.5%로 3.5 M glycerol에서 가장 높았으나 초자화동결법보다 현저하게 낮았다. 4. 토끼 상실배를 25% glycerol과 25% 1,2-propanediol을 함유한 동결액에 실온에서 10분간 노출후 초자화동결했을 때 발생율은 75.0%로 실온에서 형평한 후 초자화동결이 가능하였다. This experiment was carried out to investigate on in vitro development of rabbit monla frozen by 2-step feezing and vitrification. The results obtained from this experiment are as follows; 1. When rabbit morula in m-PBS containing 1.5, 2.0, 2.5 or 3.0 M glyceral and 0.5 M sucrose for 10 min at room temperature were cooled at -30℃ for 30 to 40 min and plunged into liquid nitrogen, the proportion of embryo developed to expanded blastoyst was 36.4, 83.3, 92.3 and 84.2%, respectively. Glycerol 2.5 M showed higher survival than others. 2. When rabbit morula in m-PBS containing 1.5, 2.0, 2.5 or 3.0 M 1, 2-propanediol and 0.5 M sucrose for 10 min at room temperature were cooled at -30℃ for 30 to 40 min and plunged into liquid nitrogen, the proportion of embrye developed to expanded blastocyst was 26.6, 55.6, 65.0 and 52.9%, respectively. 1, 2-propanediol was less effective than glycerol. 3. When rabbit morula in m-PBS containing 2.5, 3.0 or 3.5 M glycerol and 0.5 M trehalose for 10 min at room temperature were plunged into liquid nitrogen and thawed rapidly, the recovery rate of embryo was 87.5, 92.5 and 92.5%, the proportion of morphologically normal embryo was 37.5, 55.5 and 60.0%, and the proportion of embryo developed to expanded blastocyst was 13.3. 36.4 and 37.5%, respectively. The proportion of embryo developed to expanded blastocyst was higher in vitrification than in plunging into liquid nitrogen. 4. When rabbit morula were frozen by vitrification in m-PBS containing 25% glycerol. 25% 1, 2-propanediol, the proportion of embryo developed to expanded blastocyst was 75.0%, the result suggested that rabbit embryos could be frozen by vitrification after equilibration at room temperature.
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