• Journal of Embryo Transfer
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• v.9 no.3
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• pp.255-260
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• 1994

원시생식세포(primordial germ cell; PGC)는 성성숙 이후에 기능을 갖는 생식세포의 근원이 되는 세포로서, 다능성을 갖고 있는 것으로 알려져 있다. 그러므로 chimera 및 유전자 변환동물 생산을 위해 널리 사용되어 온 배아주(embrynic stem; ES)세포를 대신할 다른 세포계라고 생각되어져 많은 연구가 진행되고 있다. 본 실험은 체외배양을 통하여 원시생식세포의 증식과 확립을 위해 배양조건을 구명하고, 또한 성장인자의 효과를 검증하기 위하여 실시되었다. 원시생식세포는 12.5일째의 ICR 생쥐태아의 원시생식선 융기조직으로부터 추출하였으며, DMEM + 20% FCS + nucleosides + antibiotics로 조성된 sDMEM 배양액을 사용하여 mitomycin C로 전처리한 되먹임세포단층(feeder layer)위에서 체외배양하였다. bFGF 및 LIF를 20, 40ng/ml농도로 각각 또는 함께 첨가하여 성장인자의 효과를 검토하였다. 원시생식세포는 성에 따라 유의적인 colony 형성율을 보였고(♂:1.9 colonies / genital ridge, ♀:1.3 colonies / genital ridge), bFGF 및 LIF의 첨가 및 첨가농도에 따라서도 유의성 있는 결과를 보였다(0.3~1.9 colonies / genital riege). 그러나 3회 이상 계대배양을 할 경우, 원시생식세포의 colony를 4% prarformaldehyde로 20분간 고정한 후, tris-maleate buffer(pH 9.0)로 10분간 3회 세정하였다. Fast Red로 염색을 실시한 결과, 대부분의 colony가 염색반응을 보여 다능성을 갖는 원시생식세포의 colony임이 입증되었다. 그러나 대부분의 colony가 3회 이상의 계대배양시 생종율이 급격히 떨어지는 것을 감안하면, 또 다른 미지의 성장인자나 보다 적절한 배양조건이 요구된다고 생각된다.

• Journal of Plant Biotechnology
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• v.41 no.2
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• pp.89-93
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• 2014

Long-term subcultured rose embryogenic calluses, which had been maintained for more than 5 to 6 years since the first embryogenesis from calluses induced from in vitro roots of rose, were identified as potential material for the development of transgenic plants. The first embryogenic calluses from 'Sweet Yellow' and two breeding lines (KR056002 and KR056006) were obtained in 2007 and 2009, respectively. Subsequently, we found that plants regenerated from long-term embryogenic calluses (LEC). Whereas the LEC from 'Sweet Yellow' takes 3 to 4 months to regenerate plants, those of the two breeding lines take 4 to 5 months. This period of time is the same as that taken for plants to regenerate from the first embryogenic callus. New embryogenesis was observed from atypical bodies (ABs) that appeared during the process of long-term subculture. We found that it is possible to use the AB as a material for new embryogenesis.

• Journal of Bio-Environment Control
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• v.22 no.3
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• pp.241-247
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• 2013

For in vitro minimal-growth conservation of S. sarmentosum, the in vitro shoots with 10 mm length were cultured on Murashige and Skoog's media (MS) containing different levels of agar (0.8, 1.2, 1.6, 2%), Gelrite (0.4, 0.6, 0.8, 1%), ABA (0, 5, 10, $20mg{\cdot}L^{-1}$), and sucrose (2, 3, 6, and 9%) without subculture at $4^{\circ}C$ and $25^{\circ}C$. All media were supplemented with $0.2mg{\cdot}L^{-1}$ BA, agar and Gelrite media, with 5% sucrose, sucrose media, with 1.2% agar, and ABA media, with 5% sucrose and 1.2% agar, respectively. In vitro minimal-growth conservation in room-temperature ($25^{\circ}C$) was effective in the media containing with $10mg{\cdot}L^{-1}$ ABA or 1.6% agar, and the healthy plantlets could be preserved for 10 months without subculture. After 12 months at $4^{\circ}C$, survival rate was 100% in all media. The in vitro minimal-growth conservation in low temperature ($4^{\circ}C$) was effective in the media containing with $10mg{\cdot}L^{-1}$ ABA or 6% sucrose, and the healthy plantlets could be preserved over 18 months without subculture. Especially, long-term conservation using minimal growth of S. sarmentosum was much more efficient in the medium containing high level sucrose at $4^{\circ}C$ compared to others.

• Journal of Plant Biotechnology
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• v.32 no.2
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• pp.123-128
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• 2005

A series of experiments were performed to establish regeneration system through friable embryogenic callus (FFC) of Lilium longiflorum 'Nellie White'. Only hard and regular callus was induced from bulb scales on medium containing 2.0 mg/L dicamba and $30{\sim}90$ g/L sucrose. The induced hard callus was subcultured on medium with 2.0 mg/L dicamba and 30 g/L sucrose, and used as a material for induction of FEC. In order to induce FEC, induced hard and regular callus was chopped into $1{\sim}2\;mm$ segments, and re-cultured on medium with 2.0 mg/L dicamba and 90 g/L sucrose. FEC was induced from chopped hard calli by the subcultures of two months interval. The induction rate of FEC was enhanced when hard callus was subcultured on same medium. FEC was proliferated more than 5 times on medium with $1.0{\sim}2.0\;mg/L$ dicamba and 90 g/L sucrose. Bulblet differentiation from FEC was very favorable on MS medium supplemented with 0.1 mg/L BA, 1.0 mg/L NAA and 30 g/L maltose, but many differentiated bulblets were changed to vitrificated ones. The differentiation of normal bulblets was most effective on medium containing $0.5{\sim}1.0\%$ activated charcoal and 30 g/L sucrose.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.53-59
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• 2003

For somatic cell nuclear transfer in Hanwoo, fetal fibroblast cell lines were established from 35, 50, 70 and 90-day fetuses of Korean native cattle. The sex of these fetal fibroblast cells were analyzed by PCR using Y-specific primers and confirmed that two cell lines were female and the other two cell lines were male. Karyotyping of these cell lines indicates that the chromosome numbers of fetal fibroblast cells were not affected by passage number and more than 80% of fetal fibroblast cells have normal chromosome number. To evaluate Go stage in cell cycle of fetal fibroblast cells, Western blotting was performed to detect the expression level of PCNA which is known to be expressed in all cell cycle stages except G$_{0}$ stage. Following serum starvation or confluent culture for 7 days, fetal fibroblast cells were effectively reached to G$_{0}$ stage. The cell cycle was resumed after culture of these Go stage-fetal fibroblast cells with normal medium. These results indicates that fetal fibroblast cells originated from Hanwoo were successfully isolated and culture system and induction of cell cycle of these cells were established for somatic cell nuclear transfer in Hanwoo.woo.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.85-91
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• 1999

In sweet potato cultivars 'Mokpo #29' and 'Sanchunza', shoots from extplants were formed 100% on the MS medium with 0.1 ㎎/L NAA and 2.0 ㎎/L BA after 30 days of culture and roots produced from the base of stem at frequencies of 66.7% ('Mokpo #29') and 69.2% ('Sanchunza'), respectively, The media with 0.5∼4.0 ㎎/L BA were produced the greatest frequency of multiple shoot and the most of shoots developed rapidly into normal plantlets with rooting within 60 days of culture. Whereas the cultivar 'Keumsi' failed to produce normal shoot multiplication on the medium with cytokinins alone because of callusing of adventitious shoots. When single shoots with 1 to 2 nodes were excised from the multiple shoot or shoots covered with callus devoid of root and transferred to MS medium with 4.0 ㎎/L BA or kinetin. Host divided shoots showed the callus induction at the stem base and it was enable to obtain regenerated plantlets with shoot and root normally.

• Korean Journal of Medicinal Crop Science
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• v.2 no.1
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• pp.38-43
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• 1994

The effect of thidiazuron on callus growth and shoot regeneration of Solanumspecies was very positive. Increasing concentrations of thidiazuron stimulated callus growth of Solanum ptycanthum and Solanum nigrum. Shoot regeneration of S. ptycanthum and S. nigrum was greater on medium with thidiazuron than that with IAA and BA. Thidiazuron at $0.5{\mu}M$ or above increased the number of shoots regenerated from S. ptycanthum calli compared to the IAA with BA. High concentrations of thidiazuron $( >I{\mu}M)$ increased the number of shoots than BA or low levels of thidiazuron and IAA or BA The addition of IAA to thidiazuron media reduced S. ptycanthum shoot formation.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.43-48
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• 1997

Apical meristems of Wasabia japonica were cultured on Murashige and Skoog's medium supplemented with cytokinins alone or together with 1.0 mg/L IAA. Shoot initials could be induced from leaf primordia on apical meristems. Calli and roots were formed on the medium containing cytokinins and 1.0 mg/L IAA in combination after 30 days of culture, but there were no callus proliferation. Shoot organogenesis began after 60 days of culture and these small shoots elongated when transferred to a medium containing 1.0 mg/L BA or kinetin. Shoots were formed directly without callus induction from apical meristems all the explants on the medium containing cytokinins variously, and most of the shoots proliferated multiple shoots which could be divided to obtain plantlets. Shoot multiplication rate in response to cytokinins was best on the medium containing 1.0 mg/L BA or 2.0 mg/L zeatin. Divided plantlets rooted well on MS medium containing 0.01 mg/L IBA after 15~30 days of subculture and the rooted plantlets developed into whole plants with multiple shoots. After rooting, the regenerated plants were washed and transferred to the pots containing sterilized soil.

• Journal of Mushroom
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• v.11 no.1
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• pp.24-31
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• 2013

As a result of advenced research, Penicillium growth inhibition effect in media of cauliflower mushroom by different LED lighting color inhibited all treated groups, but blue wavelength treatment group was unfitted for culture of cauliflower mushroom due to lots of spore of penicillium. So, to investigated characteristics of mycelial growth of cauliflower mushroom according to different LED wavelength and LED wavelength color. As a results, all red wavelength treatment groups found highest mycelial growth tendency. Thus, mycelial growth investigated different quantity of red lighting wavelength conditions. The quantity of lighting wavelength was low intention, 1.41 ${\mu}mol/m^2S$ treatment group found highest mycelial growth. Effects of mycelial growth by subculture found difference of statistical in one time to carry out a subculture treatment group. Mycelial growth by different quantity of LED lighting in different media composition of wood chip media indicated highest trend in the Korean pine treatment groups. To cultured treatment group for 84th days found difference of statistical, when a quantity of LED lighting red wavelength 2.11 ${\mu}mol/m^2S$ treated in wood chip of the Korean pine media. In conclusion, good culture condition of cauliflower mushroom estimated quantity of red lighting wavelength 2.11 ${\mu}mol/m^2S$ in wood chip media of the Korean pine for 84th days.

• Journal of Ginseng Research
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• v.24 no.4
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• pp.157-161
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• 2000

Cell growth and production of phenolic compounds by callus cultures of Panax ginseng C. A. Meyer were investigated under various phytohormones concentrations and inoculum size. The results indicated that the cell growt was improved by a MS medium supplemented with 2 mg/L of CPA. The maximum cell yield was obtained at inoculum size of 1 g/flasd. The production of phenolic compounds in the callus cultures was higher than those in the ginseng root. Especially, one cell line (20601) showed the highest content of phenolic compounds and antioxidant activity.