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강만종,이철상,한용만,유대열,이경광 제주대학교 농과대학 제주도축산문제연구소 1991 畜産論叢 Vol.6 No.1
This study was carried out in order to investigate eggects of cryoprotectant concentration and equilibration time on survival of ultrarapidly frozen 2-cell mouse embryos Mouse 2-cell embryos. fol-lowing dehydration by exposure to DMSO and sucrose. were directly immersed into liquid nitrogen and thawed in 37℃ water. Viability was defined by development rate to the blastocyst stage after in vitro culture for 72 hours. The results are summarized as follows ; 1. When 0.25M of sucrose was added into the freezing medium a t various concentrations of DMSO and dilution medium, higher development rate of embryo was obtained in 3.0M DMSO conentrations (82.6%). However, When sucrose concentrations of 0.25 and 0.5M were added to the freezing medium with 3.0M DMSO and dilution medium, development rate of embryos were 81.7% and 24.1%, respectively. 2. In the equilibration time at room temperature, higher development rate was attained after short period of time(2.5min) in 3.0M DMSO +0.25M sucrose(85.9%). 3. The development rate of embryos at in vitro 2-celL in vivo 2-celL solution control and untreated control was 84.6%, 90.9%, 89.9%. and 89.7%. respectively.
Mouse 수정란의 급속동결에 관한 연구 제II보 Mouse수정란 급속동결에 있어서 수정란의 발육단계와 식빙(seeding)이 생존율에 미치는 영향
강만종,김영훈,문성호,김중규 한국동물번식학회 1989 Reproductive & developmental biology Vol.13 No.3
The effects of seeding method and optimum time for freezing embryos according to the developmental stages on embryo survival rates after rapid freezing were determined using the FDA-test. The summarized results are as follows : 1. In the rapid freezing of embryos, the sucrose added medium together with Co-seeding or non-seeding showed the FDA scores of 4.67 and 4.20, respectively, but, raffinose addition obtained FDA scores of 4.27 and 3.97. 2. The developmental stage of embryos at freezing was most critical on the survival of embryos after thawing. Higher FDA scores were obtained in the order of blastocyst stage(4.94), morula stage(3.82) and ealy stage(2.65) in sucrose added medium. The same trend was observed in the raffinose added medium with an order or 4.91, 4.47 and 2.32. 3. Microscopic study of embryo before freezing and post-thawing indicated that the embryo showed shrinkage within 5 minutes after the embryo was transfer to the freezing medium. When thawed embryo was tranfered to the dilution medium, swelling of the embryo was observed and there after it reshrank indicating the removal of cryoprotectant from the embryo. The size of the embryo recovered to the original state when it was moved into a PBS-solution.
할구 한 개가 제거된 생쥐 4세포기 수정란의 초급속동결
강만종,이철상,한용만,유대열,이경광 韓國受精卵移植學會 1992 한국동물생명공학회지 Vol.7 No.2
Cryopreservation of mouse embryos biopsied at 4-cell stage was investigated by ultrarapid freezing. Four-cell embryos were obtained from ICR mice on 55h after hCG injection. Zona pellucida of the embryos were partially dissected with a cutting pipet, and then single blastomeres were biopsied from the embryos followed by incubation in + and +-free M16 medium for 30min. Biopsied embryos cultured for lh or 15h were frozen by ultrarapid freezing method using 3M DMSO or 5M glycerol as a cryoprotectant, respectively. The developmental rate of biopsied embryos after ultrarapid freezing and thawing to blastocysts was 81 % in the group of biopsied embryos cultured for lb and 98% in the group of biopsied embryos cultured for 15h, respectively. When biopsied embryos after ultrarapid freezing and thawing were transferred to the uteri of pseudopregnant recipients, normal live young were born. These results suggest that this freezing method can efficiently cryopreserve biopsied mouse embryos.