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비특이 면역증강제 $Barodon-FX^{(R)}$ 첨가 TCM199에서 생쥐 및 소 초기배의 체외 배반포 발달에 관한 연구
정영채,나광빈,김창근,류재원,최수일,전경수,류범룡 한국동물번식학회 2001 Reproductive & developmental biology Vol.25 No.2
This experiment was designed to evaluate effects of nonspecific immunostimulator(NIS) Barodon-FX(equation omitted), anionic alkali mineral complex and far-infrared radiation solution on in vivo-produced mouse and in vitro-produced bovine embryos to blastocyst development. Proportion of mouse embryos developing into blastocyst was not greater in BSA- and Barodon-added medium than in BSA-control, but there was signifcantly different(P < 0.05) in hatching and hatched blastocyst development between 0.25% Barodon-and PVP-contained medium(54.7%) than PVP-control(32.5%). BOEC and GC resulted in higher proliferation rate(24∼40% and 17∼22%, respectively) in 0.25∼0.5% Barodon-added medium than in controls, but proliferation of GC and CC greatly decreased in 1∼2% Barodon-added medium. Effect of Barodon on cell proliferation greatly varied among somatic cells. Proportion of early bovine embryos developing into morula and blastocyst was significantly greater(P < 0.05) in 0.5% Barodon-added medium(50% and 63.6%) than in control(31.6% and 27.4%) under co-culture with BOEC and GC, but developmental rate was not different between other Barodon treatments and control. These data indicate that effect of Barodon on cell proliferation significantly varied between somatic cells and that addition of 0.5% Barodon in BOEC-coculture system may further improve blastocyst development in early bovine embryos.
전압, 공핵배 발생단계 및 체외배양액이 소의 제2세대 핵이식배 발생능에 미치는 영향
정영채,김창근,나광빈,김광식,박세영,류재원,오신애,박진서 中央大學校 遺傳工學硏究所 1998 遺傳工學硏究論集 Vol.11 No.1
소에서 체외에서 생산된 제1세대 핵이식배를 다시 공핵배로 사용하여 제2세대 핵이식배를 생산하기 위한 효과적인 방법을 개발하기 위하여 본 연구에서 세포융합시 전압, 제1세대 핵이식배의 발생단계 및 체외배양조건에 따라 제2세대 핵이식배의 발생결과를 조사한 결과는 다음과 같다. 1. 세포융합시 전압을 0.7kV/cm, 1.0kV/cm 및 1.2kV/cm로 하였을 때 세포융합율과 난할율에서 유의차는 없었으며 후기배의 발생율이 1.0kV/cm가 0.7kV/cm와 1.2kV/cm보다 다소 높게 나타났다. 2. 8-15, 16-23 및 25-32세포기의 제1세대 핵이식배를 공핵배로 이용한 제2세대 핵이식배의 융합율, 난할율 및 상실배까지의 발생율은 세포기에 따라 유의차가 없었다. 그러나 배반포배 발생율은 각각 3.4%, 6.9% 및 18.5%로서 16-23세포기와 25-32세포기간에는 유의차가 없었으나 25-32세포기는 8-15세포기보다 유의적으로 높았다(p<0.01). 3. 제2세대 핵이식배를 BOEC, BOEC-CM 및 10% FBS+BOEC-CM에서 배양한 결과 난할율과 상실배까지의 발생에서 큰 차이가 없었으나 배반포까지의 발생율은 각각 12.5%, 5.7% 및 9.6%로 BOEC 처리구에서 처리구가 가장 높은 것으로 나타났다. This study was carried out to investigate the effect of the electric pulses, donor embryo cell-stages and culture media on the 2nd generation nuclear transplant bovine embryos. Parent donor embryos were used at day 4 after IVF and 8 to 32 cell-stage embryos at day 3∼5 after the first cycle nuclear transfer were used as donor for 2nd generation. The recipient oocytes were enucleated 22 to 23 h after the onset of in vitro maturation(IVM) and preactivated 24 h after IVM. Electric fusion was performed 30 h after IVM. When a pulse of 0.7kV/cm, 1.0kV/cm or 1.2kV/cm direct current for 70μsec was applied, the fusion rates of the 2nd generation were 73.5%, 87.3% and 83.6%, respectively and the cleavage rates were 78.0%, 80.6% and 86.6%, respectively. The proportions of fusion and cleavage were lower in 0.7kV/cm than in 1.0kV/cm and 1.2kV/cm. The developmental rate to blastocyst in the 2nd generation nuclear transplant embryos was higher in 1.0kV/cm(12.9%) than in 0.7kV/cm(8.0%) and 1.2kV/cm(6.5%). The rate of fusion and cleavage and developmental rate to blastocyst in the 2nd generation embryos derived from 8 to 32 cell-stage donor embryos was 87.1%, 85.2% and 18.5%, respectively, from 25-32 cell-stage, 86.6%, 79.3% and 6.9%, respectively, from 16-23 cell-stage, and 84.1%, 77.6%, and 3.4%, respectively, from 8-15 cell-stage. These rates were not significantly different between 25-32 cell-stage and 16-23 cell-stage donor embryos, but the developmental rate was higher from 25-32 cell-stage donor embryos, than from 8-15 cell-stage. The developmental rate to blastocyst in the 2nd generation nuclear transplant embryos was higher in co-culture with bovine oviduct epithelial cells(12.5%) than in BOEC-conditioned medium with 10% FBS(9.6%) or without FBS(5.7%). The developmental rate in co-culture with BOEC was higher than in BOEC-conditioned medium.
체외수정 및 미세조작에 의한 가축배의 생산과 효율적 이용에 관한 연구 6. 소 핵이식배의 체외배양 , 동결보존 및 성판정에 관한 연구
정영채,김창근,윤종택,나광빈,오성종,이종완,김흥률,김광식,박선애,유영아 ( Y . C . Chung,C . K . Kim,J . T . Yoon,G . B . Luo,S . J . Oh,J . W . Lee,H . R . Kim,K . S . Kim,S . A . Park,Y . A . You ) 한국축산학회 1997 한국축산학회지 Vol.39 No.3
This study was carried out to establish an effective method for the improvement of subsequent development after nuclear transfer(NT) in bovine embryo, for the extension of utility through freezing and sexing of NT embryos. The rates of cleaved and developed embryos upto morula-blastocyst stages after the NT in in vitro produced embryos were 60.6 and 3.7%, respectively. Whereas, in in vivo embryos, slightly higher developments were obtained(70.8 and 5.6%), respectively. No difference was found in embryonic development upto morulablastocyst stages among the different levels of EGF added to the serum containing medium. The rates of cleaved and developed embryos upto morula-blastocyst were higher when electrostimulation was performed either before fusion or after addition at 21h IVM. The survival rate of frozen NT embryos was higher in late morula than that of earlier embryos. PCR-based sexing of NT embryo demonstrated that higher proportion of embryos was male(1.9:1), instead of normal ratio(1:1).
돼지에 있어서 분만시각의 인위적 조절에 의한 자돈생산 효율에 관한 연구 I. 돼지에 있어서 자돈분만 시각에 미치는 요인에 관한 연구
정영채,김창근,윤종택,이종완,전광주,이석우,이학철,이관순,나광빈 한국동물번식학회 2000 Reproductive & developmental biology Vol.24 No.2
To improve the pig farm management and efficiency of swine industry by inducing the farrowing to day-time from night, In the first experiment, reproduction records of purebred and crossbred pigs were collected and analyzed to estimate the $\varepsilon$ ffec 잉 of factors affecting day and night farrowing. The general linear model was used to estimate the least square means of the factors affecting various reproductive characteristics. And also, chi-square tests were used to examine the independence of the reproductive traits and environmental factors using the SAS (1992). The comparisons between pure and crossbred pigs for total number born, percentage of number born alive, gestation length, time length for farrowing were determined. The results indicated that the gestation length of crossbred (115.11 d) was slightly longer than that of purebred (114.89 d, p<0.05). For the seasonal effects on total number born, the largest was found in spring and no differences were found among summer, fall and winter. The average gestation length was 1 day longer in spring and winter than in summer and fall. The total number born and number born alive were smaller in first, second, and greater or equal to sixth parity than other parities. And also, the percentage of no. born alive was least in greater or equal to sixth parity. For the effect of mating methods, natural and artificial insemination, on total number born and number born alive, no differences between the two methods were found. However, the percentage of number born alive for natural mating was 98.06% and was higher than artificial insemination(93.75%). The time length for farrowing was I hour were found for the 6 hrs of farrowing time. In general situation of pig farms, day-time farrowing was 34.8% and night farrowing was 65.2%, indicating that night farrowing was almost double of the night farrowing.