• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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• pp.713-717
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• 1998

The synergistic effect of fetal calf serum (FCS) and chicken embryo extract (CEE) on protein synthesis of chicken embryo myoblasts was examined. Myoblasts were derived from chicken embryo cultured for 14 days by trypsin digestion and cultured in 5% $CO^2/95%$ air at $37^{\circ}C$. When myoblasts were cultured at the low level of cell density (20-50% of well), CEE enhanced the ability of FCS to stimulate protein synthesis of myoblasts. However, there was no significant effect of CEE to stimulate protein synthesis of myoblasts cultured at high level of cell density (100% of well).

• Reproductive and Developmental Biology
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• 제29권3호
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• pp.145-147
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• 2005

Autocrine or paracrine mediators released by the early embryo are implicated in the support of embryonic development. Their mechanisms and optimal embryo density in the medium, however, are uncertain. This study was conducted to establish the optimal embryo density and culture medium volume in mouse parthenogenetic embryo culture. In experiment 1, culture of parthenogenetirally activated oocytes at a concentration of $2{\~}4$ embryos/${\mu}L$ significantly improved development to the blastoryst stage ($72{\%}{\leq}$) compared with culture at the lower ($0.2{\~}1$e mbryos/${\mu}L,\;0\~37.5\%$) and the higher ($5{\~}6$ embryos/${\mu}L,\;30\~53\%$) concentration for 120 h when the oocytes were cultured in a 5 ${\mu}L$ drop under mineral oil In experiment 2, the embryos cultured at a concentration of $2{\~}4$ embryos/${\mu}L$ in a 10 ${\mu}L$ drop ($81.1{\%}$) showed significantly higher blastocyst rates than those in a 5 ${\mu}L$ drop ($68.5{\%}$). This study optimizes in vitro culture condition by modifying embryo density and the volume of culture medium It may give appropriate level of autocrine and/or paracrine factors to enhance viability and subsequent normal development of mouse parthenogenetic embryos in vitro.

• 한국수정란이식학회지
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• 제25권4호
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• pp.259-262
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• 2010

Optimization of the preimplantation mammalian embryo culture condition was widely focused on refining medium composition under the name of chemically defined media. However, recent research revealed that the alteration of physical environment can be a crucial factor to a successful embryo development. In this study, under the same embryo density, a novel culture device named oil-free micro tube culture (MTC) system was evaluated using porcine parthenogenetic embryos. The activated oocytes were placed into the 0.2 ml thin-wall flat cap PCR tube and cultured to the blastocyst stage. As a preliminary step, embryo density and culture medium volume were optimized under a standard drop culture system. The optimal embryo density range for in vitro culture was 0.5 embryos per ${\mu}l$ in $20\;{\mu}l$ drop (20.5%) and 1.0 embryos per ${\mu}l$ in $10\;{\mu}l$ drop (20.6%). Based on these results, we compared drop culture system and 'MTC' system in terms of the developmental rate to the blastocyst stage. In $20\;{\mu}l$ medium volume, the 'MTC' system showed similar blastocyst formation rate when compared with drop culture system (20.2% versus 20.5%, respectively) while the 'MTC' system showed lower blastocyst formation rate than drop culture system in $10\;{\mu}l$ one (12.7% versus 20.0%, respectively). Therefore the $20\;{\mu}l$ MTC system may be an alternative incubation system for short-distance embryo transport without carrying the $CO_2$ incubator and this provides novel embryo culture device to clinical veterinary embryologists.

• 한국작물학회지
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• 제37권2호
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• pp.134-140
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• 1992

당근종자의 발아율의 낮음과 발아지정의 문제점을 해결하기 위해서 Danver 126품종을 공시하여 여러가지 prime처리와 density separation 방법을 이용한 결과 다음과 같은 효과를 얻었다. 1. 여러가지 prime처리중 SMP처리(수분함량 90%, 6일간처리)에서 배의 생장이 아주 양호하였고 발아속도 및 발아율이 가장 좋았으며 종자에 피해도 없었다. 2. 무기염처리(0.2M KNO$_3$＋0.1M $K_2$HPO$_4$)에 서는 배의 생장은 좋았으나 염에 의해 종자가 피해를 입었다. 3. SMP처리(수분함량 90%, 6일간)된 종자를 density separation한 결과 종자의 비중이 낮을수록 배의 생장이 컸으며 발아율 및 발아속도가 빨랐다.

• 한국수정란이식학회지
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• 제15권2호
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• pp.129-136
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• 2000

This study was carried out to investigate the effect of biological factors on the in vitro production(IVP) of bovine oocytes for development of simple culture methods and medium. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol and this study was examined if there were necessary to co-culture, media change, media type and embryo density. This results were as follows: 1. The development rate according to co-culture with cumulus cells and non co-culture as drop culture was not significantly different in cleavage (68.9 vs 71.7%), 8-cell stage (41.2 vs 44.1%) and blastocyst stage (12.2 vs 13.8%), respectively (p<0.05) 2. The blastocyst development rates in YS and CRIaa were higher than that in TCM199 (12.4, 10.4$ vs 3.7%), but the cleavage (69.0, 77.8 and 61.0%) and 8-cell stage (31.7, 37.0 and 35.7%) development accoring to YS, TCM199 and CRIaa ws not significantly different, respectively (p<0.05). 3. There was no significantly different in cleavage (62.6, 59.5 and 61.2%), 8-cell(34.7, 37.9 and 34.0%) and blastocyst (9.5, 11.6 and 12.8%) development among medium change time as control, Group I and Group II, respectively (p<0.05). 4. Blastocyst formation of 8-cell stage according to embryo density was not significantly different in 1, 10 and 25 embryos (27.3, 22.5 and 34.0%), respectively (p<0.05). These results indicated that simple culture system could reduce bovine IVP embryos as drop culture as non co-culture system, high density embryo (25 embryos/50 $\mu$1 drop). YS defined medium and no medium change for whole culture period, although other biological factors need to examine in order to produce efficient IVP bovine embryos.

• 한국수정란이식학회지
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• 제15권3호
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• pp.271-277
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• 2000

This study was conducted to establish the optimal culture conditions for in vitro production of bovine embryos derived from slaughter house ovaries. Cumulus-oocyte- complexes (COCs) collected by aspiration from follicles of 2~7 mm in diameter were matured in Ham's F-10 medium supplemented with 0.01 $\mu\textrm{g}$/m1 epidermal growth factor (EGF) at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. After 24 hrs of culture, the oocytes were co-cultured with epididymal sperm selected off by Percoll-density gradient in TALP medium for 24 hrs. The presumptive zygotes were cultured in HECM-6 medium for 3 d post-insemination, and followed by cultured in TCM199 medium until 7 to 10d post-insemination. The cultures were compared of their cleavage and development into later stage in culture medium by additions of different protein sources (PVA, BSA and BCS) and by different embryo density. The rates of cleavage and development rates into blastocyst were not significantly (P<0.05) different among the culture media containing with BSA (75.0% and 40.5%), BCS (76.7% and 38.0%) and PVA (72.5% and 42.2%), respectively. Significantly (P<0.05) higher blastocysts rates were obtained in culturing of 30 and 40 embryos in each 50$\mu$l droplets of culture medium than in 5, 10 and 20 embryos. These results indicate that the optimal density of embryos is 30~40 embryos in a 50$\mu$l droplet of culture medium. Furthermore there is no effect of different protein sources on early embryonic development.

• Applied Biological Chemistry
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• 제51권3호
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• pp.200-206
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• 2008

본 연구는 쌀 배아를 활용한 만성 대사성질환 예방용 제품 및 건강기능성 식품 개발의 일환으로써 고콜레스테롤 식이를 급여하여 고지혈증을 유발시킨 흰쥐에서 15%의 배아와 25%의 흑미 미강색소 배아젤리의 첨가가 혈장과 간 조직의 지질 대사와 항산화효소의 환성에 미치는 영향을 살펴보았다. 실험 식이를 6주 간 급여한 결과, 배아와 흑미 미강색소 배아젤리 첨가는 실험동물의 식이섭취에 영향을 미치지 않았다. 배아군과 흑미 미강색소 배아젤리군은 고콜레스테롤 급여 대조군에 비해 혈장의 총 콜레스테롤과 LDL-콜레스테롤 및 간의 중성지방과 총 콜레스테롤 농도를 감소시키고, HDL-콜레스테롤 농도와 HDL-C/TC 비는 증가시켰으며, 동맥경화지수는 감소시켜 체내 지질대사의 개선 효과가 있었다. 혈장 GOT와 GPT 수치는 배아와 흑미 미강색소 배아젤리를 첨가하였을 때 감소하여 고콜레스테롤혈증 상태에서 간 기능 보호에 긍정적인 효과가 있었다. 또한 배아와 흑미 미강색소 배아젤리는 고콜레스테롤 급여로 인해 증가된 혈장과 간 내의 지질과산화를 억제시키는 효과가 있었다. 반면, 항산화 효소인 간 조직의 SOD와 CAT활성은 배아와 배아젤리 첨가에 따라 증가하였다. 이상의 결과로 볼 때 배아와 흑미 미강색소 배아젤리는 고콜레스테롤 식이 흰쥐의 간 조직에서의 항산화 활성을 강화시키고 산화적 손상을 억제시키는 자용이 있으며 현장과 간조직의 지질대사를 개선하여 심혈관계 질환을 예방 및 감소시킬 수 있을 것으로 사료된다.

• Journal of Plant Biotechnology
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• 제49권1호
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• pp.61-73
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• 2022

In this study, we developed a reliable and efficient Agrobacterium-mediated genetic transformation system by applying sonication and vacuum infiltration to six chickpea cultivars (ICCV2, ICCV10, ICCV92944, ICCV37, JAKI9218, and JG11) using embryo axis explants. Wounded explants were precultured for 3 days in shoot induction medium (SIM) before sonication and vacuum infiltration with an Agrobacterium suspension and co-cultivated for 3 days in co-cultivation medium containing 100 µM/l of acetosyringone and 200 mg/l of L-cysteine. Responsive explants with putatively transformed shoots were selected using a gradual increase in kanamycin from 25 mg/l to 100 mg/l in selection medium to eliminate escapes. Results showed optimal transformation efficiency at a bacterial density of 1.0, an optical density at 600 nm wavelength (OD₆₀₀), and an infection duration of 30 min. The presence and stable integration of the β-glucuronidase (gusA) gene into the chickpea genome were confirmed using GUS histochemical assay and polymerase chain reaction. A high transformation efficiency was achieved among the different factors tested using embryo axis explants of cv. JAKI 9218. Of the six chickpea cultivars tested, JAKI9218 showed the highest transformation efficiency of 8.6%, followed by JG11 (7.2%), ICCV92944 (6.8%), ICCV37 (5.4%), ICCV2 (4.8%), and ICCV10 (4.6%). These findings showed that the Agrobacterium-mediated genetic transformation system will help transfer novel candidate genes into chickpea.

• Journal of Plant Biology
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• 제35권1호
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• pp.45-51
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• 1992

인삼(Panax ginseng C.A. Meyer)의 채종전 홍숙 종자로부터 채종 후 미개갑 종자까지 단계별 배발달에 따른 배유세포내 단백과립의 변화상을 확인하여 다음과 같은 결과를 얻었다. 홍숙 초기단계의 종자의 배유세포에는 구형의 스페로솜이 산재하였으며, 액포내에는 저장 단백질이 축적되어 단백과립을 형성하였다. 홍숙 말기단계의 종자의 배유세포내 세포질은 대부분 스페로솜과 단백과립으로 충만하였고, 세포소기관은 거의 관찰할 수 없었으며, 단백과립은 전자밀도가 높은 무정형의 함유물을 가지고 있는 것과 균일한 단백질 기질로만 이루어져 있는 것 등으로 크게 구분되었다. 채종 후 후숙 처리를 하지 않은 미개갑 종자에서, 배유세포내 단백과립은 구상체(globoid), 단백질 결정체 등을 함유하고 있었고, 구상체는 다양한 형태의 전자밀도가 높은 물질을 가지고 있었다. 제형층은 배와 배유조직 사이에 위치하였으며, 이와 인접하여 배유세포벽의 분해양상과 셀루로우즈 미세섬유상이 관찰되었다. 제형층은 lipid body와 분해된 배유세포의 잔유물로 이루어져 있었다. 제형층과 인접한 배유세포의 단백과립은 퇴행성 변화를 나타내었고, 이로 인해 단백질 기질은 전자밀도가 점진적으로 낮아지는 결과를 얻었다.

• Environmental Analysis Health and Toxicology
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• 제22권3호
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• pp.211-218
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• 2007

Even though some standard developmental bioassay protocols for environmental assessment using sea urchins have already been described, there have not been many attempts to apply and modify these protocols with Korean species. Therefore, there is a strong need to establish standard bioassay protocols using sea urchins commonly found in Korea. Prior to developing a new protocol, it is essential to know the optimal conditions for the bioassay procedures. We investigated the optimal conditions (temperature, salinity, and embryo density) of the sea urchin Strongylocentrotus intermedius. The ideal temperature for developmental bioassay of S. intermedius was determined to be $15^{\circ}C$ and the time required for the embryo to become pluteus larva was 72 hr. The optimal range of salinity for the embryo toxicity using S. intermedius was between 30 to 32 psu, which is similar to the range found in the natural habitats of adult populations. The optimum density of embryos at the beginning of bioassays was 100 embryos/mL. When the assays were carried out at higher densities, the proportion of normally developed larvae decreased significantly.