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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
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.