• 한국수정란이식학회지
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• 제20권2호
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• pp.123-128
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• 2005

본 연구는 돼지 난포란의 vitrification 동결 시 내동제의 종류 및 농도가 생존율에 미치는 영향과 수정 후 체외발생율을 조사하고자 수행하였다. 1.0, 15, 30 및 40시간 성숙 배양시킨 난포란을 vitirfication 동결보존 후의 MII로의 발생율은 각각 $56.7\%,\;53.3\%,\; 63.3\%,\;65.0\%$였으며, diploid로의 발생율은 $23.3\%,\;18.3\%,\;10.0\%,\;3.3\%$로서 대조군의 ME 단계의 $78.2\%$에 비해 낮게 나타났으며 diploid 단계의 $5.5\%$에 비해서는 높은 체외성숙율을 나타냈다. 체외발생율은 초기의 미숙 난포란일수록 높은 체외성숙율을 나타냈다. 2. 미성숙 난포란을 회수 후 0, 15, 30 및 40시간 성숙 배양시킨 난포란을 vitirfication 동결 후 융해하였을 때 생존율은 각각 $34.0\%,\;26.0\%,\;18.0\%,\; 12.0\%,\;10.0\%$로서 비동결 난포란의 $60.0\%$에 비해 낮게 나타났지만 비교적 양호한 생존율을 나타냈다. 3. 0, 15, 30 및 40시간 성숙 배양시킨 미성숙 난포란을 vitrification 동결 융해 후 수정시켰을 때 체외수정율은 $60.0\%,\;54.0\%,\; 48.0\%,\;38.0\%$였으며, 배반포로의 체외발생율은 각각 $26.0\%,\;18.0\%,\; 8.0\%,\;4.0\%$로서 비동결 대조군의 $78.0\%$와 $38.0\%$에 비해 낮은 체외수정율과 체외발생율을 나타냈다. 4. 돼지 난포란을 EDS와 ETS 액으로 vitrifiestion 동결운해 후 $0\~15$ 시간 배양한 다음 체외수정시켰을 때 체외 수정율과 발생율은 각각 $50.0\%,\;22.0\%$ 및 $46.0\%,\;18.0\%$로서 대조군의 $74.0\%$와 $38.0\%$에 비해 낮게 나타났다.

• 한국가축번식학회지
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• 제22권2호
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• pp.187-194
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• 1998

본 연구는 돼지 수정란의 동결에 있어서 내동제의 종류, sucrose 농도, 평형시간 및 융해온도가 생존율에 미치는 영향과 미성숙 난자의 체외배양과 동결융해 후 생존율과 체외수정율을 조사하고저 수행하였다. 1. 수정란의 급속동결에 있어서 1.5 M glycerol, 2.0 M DMSO, 2.5 M ethyleneglycol 및 2.0 M propanediol 농도에서 생존율이 유의하게 높았으며, 내동제에 첨가된 sucrose농도는 0.25 M 첨가가 다른 농도에 비해 생존율이 높게 나타났다. 동결수정란은 3$0^{\circ}C$에서 융해시 생존율은 33.3~40.6%로서 $25^{\circ}C$ 및 37$^{\circ}C$의 융해온도에 비해 높게 나타났으며, 동결시 1.5 M 및 2.0 M glycerol＋0.25 M sucrose가 첨가된 내동액으로 짧은 평형시간(2.0~5.0분)이 긴 평형시간 (10~15분)보다 높은 생존율을 나타냈다. 2. 미성숙란을 1~8시간 체외배양시킨 다음 동결융해 후 체외수정시켰을 때 수정율은 6.7~26.7%로서 단시간의 체외성숙 배양이 높게 나타났으며, 또한 체외배양시 생존율은 10.0~30.0%를 나타냈다.

• 한국가축번식학회지
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• 제18권4호
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• pp.251-256
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• 1995

Two experiments were conducted to study the production of in vitro fertilized bovine embryos and the viability of blastocysts cryopreserved by vitrification. In experiment 1, production rate of in vitro matured bovine oocytes after fertilization in medium containing bovine oviduct epithelial cells (BOEC), cumulus cells and granulosa cells to blastocysts were 18.4, 14.6 and 13.1%, respectively. Developmental percentages of blastocysts produced at day 6, 7 and 8 were 8.5, 10.6 and 15.2% respectively. Hatching rate of bovine embryos produced was 60.0%. In experiment 2, post-thawed surviving embryos in a vitrification solution consisting of 7.15M ethylene glycol, 2.5 mM ficoll and 0.3 M sucrose were 36.4% (56/154). Also, survival rate of bovine embryos after exposed to vitrification solution at 1, 2, 3, 4 and 5 min were 84.0, 88.0, 71.0, 48.0 and 24.0% respectively.

• 한국어병학회지
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• 제20권3호
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• pp.257-267
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• 2007

The hypo-salinity effects on fertilized eggs, embryos and larvae were investigated in olive flounder (Paralichthys olivaceus) obtained from hatcheries in Cheju-Island, Yeosu and Chungnam. Those were treated to eight concentration; 0, 3.4, 6.7, 10.1, 13.4, 20.2, 27.4 and 33.6 ‰. It was not discrepancy in the survival rate and hatching success rate of fertilized embryos obtained from different regions. Also, in the larvae, the regional difference was not appeared. The survival rate and hatching ability of embryos significantly diminished in the lower groups than 13.4 ‰ compared to 33.6 ‰. After fertilization, namely embryos are tolerant of a wide range of salinity (13.4 - 33.6 ‰). Reduced salinity induced an increase of the malformed embryo and larvae including various deformities; irregular embryos membrane (or yolk sac depression), fin erosion and swim bladder inflation in the flounder embryo. The hatching success of embryos was significantly reduced in lower salinity than 13.4 ‰. Notably, the reduction of larval survival rate significantly was observed in ≤10.1 ‰ treated groups with the same manner of survival rates of the embryos. Additionally, olive flounder was found to be adequate model for measuring external impulses because there are no the regional differences.

• 한국산림과학회지
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• 제103권4호
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• pp.703-708
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• 2014

본 연구는 몽골 반건조 지역 현지에서 토양 개량이 묘목의 활착 및 생장에 미치는 효과를 알아보기 위해 수행되었다. 이를 위하여 알칼리성 사질 토양인 연구대상지에 2년생 백양나무(Populus sibirica)와 비술나무(Ulmus pumila) 묘목을 심고 질소(시비량에 따라 2수준), 황, 인공보습제, 황토가공소재 등을 처리하였다. 4개월 뒤, 두 수종의 묘목 활착률은 질소 시비량의 증가에 따라 감소하였다. 일반적으로 질소의 시비는 양분을 공급하여 묘목의 활착률 및 생장을 높이는 것으로 알려져 있다. 그러나 본 연구 결과에서 나타난 활착률 감소는 과다시비로 인한 토양 삼투압의 증가에 의한 것으로 보인다. 두 수종 모두 황토가공물질 처리구에서 근원경 생장에 유의한 증가가 나타났다. 그리고 백양나무의 경우 인공보습제 처리구에서도 근원경 생장이 유의하게 증가하였는데, 이는 백양나무가 건조에 내성이 있음에도 불구하고 수분 요구도가 높은 포플러속에 속하기 때문인 것으로 보인다. 반면 황 처리는 두 수종의 활착률과 근원경 생장 모두에서 효과가 없는 경향이 나타났는데, 이는 건조지 토양의 낮은 황산화율 때문으로 추정된다. 향후 건조/반건조 지역에서 토양 개량의 장기적인 효과를 알아보기 위하여 묘목 생장 및 토양 특성에 대한 지속적인 모니터링이 요구되며, 연구 수종에 적합한 시비량 산정을 위한 추가 연구가 필요한 것으로 판단된다.

• 생태와환경
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• 제53권3호
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• pp.221-228
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• 2020

멸종위기 양서류 금개구리의 생물학적 특성 및 증식을 위한 기초자료 확보를 위해 난 발생 및 올챙이 초기 먹이 종류에 따른 영향에 대한 연구를 수행하였다. 사육수조 내에서 자연산란한 금개구리의 수정란을 현미경 하에서 관찰하였다. 수정란은 교질층에 둘러싸인 구형으로, 동물극과 식물극이 명확하게 구분되었다. 수온 27.0±1.0℃에서 수정 후 31시간 뒤에 부화하였으며, 수정 후 175시간 뒤에 아가미 뚜껑을 완성하였다. 개구 이후 단계부터 35일간 먹이종류별 성장 및 생존율을 분석한 결과 식물성 사료와 테트라민 단독 공급군의 평균 증체율(각각 5843.9%, 5736.3%), 평균 일일성장률 (각각 11.67%, 11.62%)이 다른 공급군들 (증체율: 641.8~4625.2%, 일일성장률: 5.72~11.01%)에 비해 가장 높았고(P<0.05), 야채 공급군이 가장 낮았다(P<0.05). 평균 사료 효율은 식물성 사료공급군이 177.83%로 야채 공급군 10.29%에 비해 약 17배 높았다. 또한 식물성 사료와 테트라민 단독 공급군의 평균생존율은 각각 97.5%, 100%로 가장 높았으며, 야채 공급군은 32.5%로 가장 낮았다. 따라서 금개구리의 성공적인 증식을 위한 올챙이 먹이는 식물성 사료 또는 테트라민을 단일로 공급하는 것이 가장 효율적인 것으로 판단된다.

• 수산해양교육연구
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• 제27권4호
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• pp.981-989
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• 2015

This study aims to investigate how ovulation induction of sevenband grouper, Epinephelus septemfasciatus varies when treated with different hormonal substances such as ovaprim, pimozide, LHRHa, and HCG. As for LHRHa, we injected it in different concentrations: $50{\mu}g/kg$, $100{\mu}g/kg$, $150{\mu}g/kg$, and $200{\mu}g/kg$. All hormonal substances including LHRHa were injected into back muscles. As a result, ovulation occurred in all sample groups. The sample group injected with HCG released the largest amount of eggs, and the dimeter of eggs and oil globules were also significantly bigger than those of others. Fertilization rate, embryonic survival rate, and hatching rate were the highest in a LHRHa ($100{\mu}g/kg$) +pimozide ($1,000{\mu}g/kg$) group and buoyant rate in a LHRHa group. Ovulation rate was 100% when LHRHa was injected in different concentrations. The largest amount of eggs was released in $200{\mu}g/kg$. The diameter of eggs and oil globules were the biggest in $200{\mu}g/kg$ and the smallest in $50{\mu}g/kg$. Fertilization rate showed no significant difference through all different levels of concentration: 94.6-95.3%. Buoyant rate was the highest in $100{\mu}g/kg$ and embryonic survival rate and hatching rate in $150{\mu}g/kg$. Based on these experimental results, we concluded that LHRHa in $100{\mu}g/kg$ is the most efficient for the ovulation induction of sevenband grouper.

• 한국수정란이식학회지
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• 제29권1호
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• pp.13-19
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• 2014

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in $38.5^{\circ}C$, 5% $CO_2$ incubator. For freezing, Day 7~9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to $-7^{\circ}C$, and the straw was seeded during 8 minutes-holding time, and was cooled to $-35^{\circ}C$ at the cooling rate of $0.3^{\circ}C/min$, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method ($56.86{\pm}26.53%$) were slightly higher than those by the slow-rate freezing method ($55.07{\pm}26.43%$) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were $72.65{\pm}18.3%$ and $79.06{\pm}17.8%$, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were $66.22{\pm}18.8%$ and $45.76{\pm}12.8%$, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

• 한국수정란이식학회지
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• 제31권4호
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• pp.367-374
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• 2016

This study was conducted to evaluate the effect of transfer temperature of epididymis on survival rate of semen and development ability of B6D2F1 mice embryos. No significant differences were noted in the survival rate of semen ($59.0%{\pm}0.1$ vs. $47.6%{\pm}0.1$), in vitro fertilization rate ($90.7%{\pm}0.1$ vs. $90.7%{\pm}0.1$), developmental rate ($90.0%{\pm}0.1$ vs. $90.0%{\pm}0.1$), and blastocysts formation rate ($53.1%{\pm}0.2$ vs. $52.3%{\pm}0.2$) between groups. (NS; P>0.05). However, the zona hatched rate was significantly higher in the $4^{\circ}C$ group compared to those of the $37^{\circ}C$ group ($47.8%{\pm}0.1$ vs. $25.6%{\pm}0.2$; p<0.05). When it comes to cell numbers of blastocysts, the % ICM (/total cells) was significantly higher in the group of $4^{\circ}C$ compared to the $37^{\circ}C$ ($27.0%{\pm}0.1$ vs. $18.3%{\pm}0.1$; p<0.05). However there were no differences in total cell numbers ($72.7{\pm}31.6$ vs. $62.0{\pm}36.6$), ICM cell numbers ($17.0{\pm}7.8$ vs. $14.6{\pm}8.6$), TE cell numbers ($55.8{\pm}29.8$ vs. $64.0{\pm}24.4$), the ratio of ICM:TE ($1:4.2{\pm}4.1$ vs. $1:6.4{\pm}7.2$) between two groups (NS; P>0.05). Taken altogether, it is expected to achieve the best developmental ability of B6D2F1 mice embryos in the transfer temperature of epididymis. Also these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice. In future, therefore, it is expected that results herein might be applied for in vitro culture of human embryos.

• 한국가축번식학회지
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• 제19권4호
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• pp.259-264
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• 1996

This study was carried out to investigate on the survival rates and in vitro developmental rates of bisected bovine embryos were by manipulator and micropipette. Bisected embryos were co-cultured in 20% FCS(v/v)＋TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 72 hrs after bisection. Survival rate and in vitro fertilization rate were defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro developmental rate of biseciton embryos co-cultured in 20% FCS＋TCM-199 medium containing PMSG, hCG, PMSG＋hCG, hCG＋$\beta$-estradiol 0 to 20 hrs and 20 to 40 hrs were 36.7, 26.7, 33.3, 40.0, and 30.0, and 30.0, 33.3, 30.0, 26.7, and 26.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing hormones was significantly higher than that of non co-culture(25.0%). 2. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS＋TCM-199 medium containing oviductal epithelial cells 4 to 5 hrs and 20 to 24 hrs were 40.0 and 33.3%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing oviductal epithelial cells was significantly higher than that of non co-culture(25.0%). 3. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS＋TCM-199 medium containing cumulus cells 4 to 5 hrs and 20 to 24 hrs were 43.3 and 36.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing cumulus cells was significantly higher than that of non co-culture (25.0%).