The aim of this study was to evaluate the effects of different straw volume (0.5 and 5.0 ㎖) on cryopreservation in boar semen. Semen quality was evaluated the motility, viability, abnormality, acrosome intactness and membrane integrity. And there were also examined the development rates of IVM/IVF embryos using frozen-thawed boar semen. Boar semen were frozen until 5℃ for 3 hours using cell freezer and making the straws, and then freezing by lowing the straws into styrofoam box on the 8 cm above the LN2. In different straw volume (0.5 and 5.0 ㎖), sperm viability and abnormality were not differ in 0.5 and 5.0 ㎖ straws, but sperm motility were significantly higher in 0.5 ㎖ straws (61.3%) than in 5.0 ㎖ straws (56.3%) (p<0.05). In the Coomassie Brillient Blue (CBB), Hoechst 33258/Propidium Iodide (H258/PI) staining and Hypoosmotic Swelling Test (HOST), the acrosome intactness and sperm membrane integrity were not differ in 0.5 and 5.0 ㎖ straws, but sperm survival rate was significantly higher in 0.5 ㎖ straw (65.0%) than in 5.0 ㎖l straw (55.0%) (p<0.05). Employing the Chlortetracycline (CTC)/Hoechst33258 (H258), all treatment group were not differ in characteristic of uncapacitated acrosome-intact sperm (F), capacitated and acrosome-intact sperm (B-type) and acrosomereaction seprm (AR-type). In the developmental rate of IVM/IVF embryos using frozen-thawed boar semen in different straw volumes, the developmental rate of morula plus blastocysts were 19.9% in 0.5 ㎖ straw and 18.8% in 5.0 ㎖ straw, respectively. These results indicated that straw volume affects the semen motility and sperm survival rate, but not other semen characteristic and developmental rate of IVM/IVF embryos.

멸종위험이 큰 우리나라 재래돼지를 유전자원으로서 안전하게 보존하고 유전적 다양성을 유지하기 위한 수단으로 수정란을 채취하여 동결보존하기 위해서 미경산 재래돼지에서 과배란 유기를 위한 적정 호르몬의 수준과 수정란의 회수 및 동결보존 방법을 확립하고자 수행한 결과는 다음과 같다. 1. hCG 500IU와 PMSG를 500, 750, 1,000IU 및 hCG 750IU와 PMSG 1,000IU를 각각 투여한 재래돼지의 배란황체와 미배란난포의 수는 각각 12.

본 실험은 실험견의 정액 동결 시 희석 액에 첨가되는 Glycerol 농도, 동결속도, 동결보존액 첨가후 평형시간, 융해온도와 시간에 따라 정자의 생존성과 운동성을 조사하여 최적의 동결조건을 확립하기 위해 실시하였다. 1. 각기 다른 Glycerol 농도를 함유한 동결보존액에서 동결보존 후 융해하였을 때 4%의 Gly-cerol 농도에서 각각 68.87.4%, 73.28.3%로서 다른 군보다 유의하게 높은 생존율과 운동성 나타냈다(P<0.05). 2.

한우의 유전자원을 보존하기 위하여 정액동결 시험을 수행하였으며 고급육계통은 육질에 대한 육종가 상위 10% 이내의 자손에서 선발하였고 다 유계통은 어미의 이유시 체중에 대한 육종가 상위 10%이내의 자손에서 선발하였으며 2개 계통에서 종모우 총 13두를 선발하여 공시하였다. 정액채취는 인공질법으로 실시하였고 의빈대에 수소를 계류하고 채정대상우를 승가시켰으며, 3회 가승가후에 채정하였다. 채정후 10분 이내에 실험실로 옮겨와 검사항목을 조사하고 37에서

정자의 동결보존을 위한 새로운 기술개발 목적은 동결과정에서 최소한의 손상으로, 응해 후 최대한 높은 활력도의 정자를 얻는 것이다 정자가 난자와 수정하기 위해서는 적당한 생존성과 운동성을 유지해야 하는데, 가장 일반적인 방법으로는 정자의 진진 운동성과 첨체의 정상 여부 및 형태 검사방법 등이 있다 본 연구는 사람 정액을 동결보존 할 때 semi-programmable freezer를 이용한 완만동결 방법과, 액체질소의 vapor를 이용한 급속동결 방법이

This study was performed to evaluate whether vitrification method using ethyle glycol and eletron microscopic (EM) grid could be used far the cryopreservation of human oocytes in ART program. Surplus oocytes were obtained from consented IVF patients. These surplus human oocytes were frozen with our vitrification method, Oocytes were exposed to 1.5M ethylene glycol (EG) in DPBS far 2,5 minutes, followed by 5.5M EG plus 1.0M Sucrose in DPBS for 20 seconds. Then oocytes were transferred onto the EM grid and the grid was plunged into LN2 for storage. For thawing, oocytes containing EM grid were sequentially transferred in 1.0M, 0.5M, 0.25M, 0.125M and 0 M sucrose in DPBS solution at the intervals of 2.5 minutes. Thawed and survived oocytes were provided for ICSI. Embryos from vitrified oocytes were transferred to uterus of the patient on 4 to 5 days after ovulation in natural cycles of on 15 to 17 day of hormone replacement cycles. A total of 370 oocytes from 26 patients were thawed and 159 (43.0%) of them survived. One hundred thirty four oocytes (84.3%) were fertilized normally and 126 pre-embryos were transferred to 26 patients, resulting in 5 clinical pregnancies. The pregnancy rate per transfer was 19.2% and implantation rate was 4.0%. Among the five pregnant, 4 patients delivered 4 healthy babies and the one patient was 32-week ongoing pregnancy. From this results, vitrification using ethylene glycol as cryoprotectant and EM grid is a rapid and simple method that can be effectively applied for the cryopreservation of human oocytes in ART program.

개의 인공수정에 사용할 정자의 보존방법을 확립하기 위하여, 동결속도와 응해 온도를 설정하여 적절한 동결방법을 정립하고자 본 실험을 실시하여 다음과 같은 결과를 얻었다. 동결의 방법에 있어서는 액체질소의 표면으로부터 17 cm 높이에서 동결하는 -3/min의 동결속도로 실시하여 37에서 2분간 응해하는 방법이 가장 좋은 결과를 보였다. 생존성과 운동성에 있어서의 차이는 없지만 첨체의 intact한 비율은 약간 낮은 결과를 보였으며, 이의 보완을 위해, 액

This experiment was carried out to investigate the effects of osmolarity of thawing diluents, seminal plasma added in thawing diluents on the sperm viability and the effects of thawing temperature, the temparature of the thawing diluents on the sperm viability and acrosomal morphology of boar spermatozoa by the straw method. The result obtained were summarized as follows: 1. The sperm viablilty after thawing of the frozen semen was shown greater in the high osmolarity(392~492mOsm) than low osmolarity(300mOsm) in thawing diluent. The added levels of seminal plasma in thawing diluent did not affect the viability of frozen-thawed boar semen. 2. In terms of thawing temperature, the sperm viability was shown higher in the frozen semen thawed at 5 for one min. (p<0.01) than those thawed at 2 or 37 for one min. The sperm viability was not significant at the diluent temparature of 2or 37 after thawing: but the sperm viability was higher in thawing diluent at 2 than in that at 37. However, the effects of thawing temperature and diluent solution on normal acrosomal rate were not significant. 3. Cleavage rates of oocytes fertilized with frozen semen were 46.4% and 43.3%, respectively, which were thawed at 5 for one min. and then diluted in mBTS medium at 2or 37. To sum up, the sperm viability was shown greater at the high of thawing diluents of frozen boar semen. In terms of thawing conditions, the sperm viability was shown greater, when semen was thawed at a high temperature for a short time and then diluted at the same temperature as that in the straw.

The objective of this study was to optimize the selection of sperm sources, optimal culture systems and vitrification method depends on sperm sources. The oocytes were inseminated with either KPN 105, 114, 191, SNU 101, 102, 103 or epididymis and then embryos inseminated were cultured in oviductal cell co-culture or HECM-6 as defined me dium. The blastocysts produced were pooled according to sperm sources as KPN, SNU or epididymis and then vitrified by OPP vitrification method. The results obtained were as follows: 1. The cleavage(86.2 or 84.7%) and development rates to blastocyst (30.6 or 32.0%) were not significantly different between oviductal cell co-culture or HECM-6 culture systems(P<0.05). 2. To determine the optimal sperm sources for using IVF in this system, cleavage rates in KPN 191 and SNU 101 (74.2, 55.8%) were significantly lower rather than those in KPN 105, 114, SNU 102, 103 or epididymis (86.7, 85.1, 89.8, 85.5 or 81.2%), but development rates to blastocyst in KPN 114, SNU 103 or epididymis sperm (30.0, 33.0 or 28.6%) were significantly higher rater than those in KPN 105, 191, SNU 101, 102(21.4, 15.4, 14.9 or 25.4%), respectively (P<0.05). 3. The blastocysts produced were pooled according to sperm sources as KPN, SNU or epididymis and then vitrified by OPP vitrification method. The survival rates were not significantly different among sperm sources (89.6%: 43/48 ; 90.1%: 46/51 ; 83.3% : 20/24). These results obtained indicate that the defined medium, HECM-6, could be use to produce of IVP bovine embryos. Since the frozen semen must be required to maintain of unvariation data in IVP embryo production system, KPN 114 and SNU 103 produced in our laboratory were useful for this purpose. The blastocysts produced by different sperm sources as KPN, SNU or epididymis were vitrified by OPP vitrification method and survived very high rates. The OPP vitrification method could be susceptibility to use of IVP bovine blastocyst embryos.