• Journal of the korean academy of Pediatric Dentistry
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• v.45 no.4
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• pp.492-498
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• 2018

This study was conducted to investigate the characteristics of subculture times in the early, middle, and late passages by measuring the time under subculture until it was judged that the supernumerary tooth-derived pulp stem cells (sDPSCS) were no longer proliferating. Three supernumerary teeth from two healthy six-years old boys were extracted and stem cells were obtained from the pulp tissue. This was called SNT1 (supernumerary tooth 1), SNT2, and the supernumerary tooth from another child was named SNT3. SNT1 and 2 were subcultured at the same time and SNT3 was subcultured a little faster. The mean time of complete subculture was $3.6{\pm}1.1$ days. Total passages were cultured up to $23.3{\pm}0.6$ and took 83 days. These were divided into three groups based on the passage. The increase rate of time taken in subculture between group I and group II was 11.9%, but the rate between group II and group III was 28.6%, which was 2.4 times increased. The time taken between passages during long-term subculture up to 22 passages shows a regressive pattern y = 0.1169x + 2.25 and y = 0.1169x + 2.0. In conclusion, the passage time of SPSCs increased in late passages, and it shows a similar pattern.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.2
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• pp.225-232
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• 1997

폐색성 혹은 비폐색성 무정자증에서 부정소 정자채취법 등이 부적절하다고 여겨질때는 정소 조직을 일부 절제하여 그 조직으로부터 정자를 직접 채취하게 되는데 일반적으로 이렇게 정소로부터 추출한 정소정자는 운동성이 전혀 없거나 매우 약한 운동성을 보이는 경우가 많다. 본 연구의 목적은 이러한 정소정자를 Vero cell과 공배양을 시킴으로써 운동성을 획득시키거나 향상시키고 이를 수정시키는 시기까지 지속시킴으로써 정소정자추출술 (TESE)을 시행하는 환자나 의료진들에게 보다 편안하고 융통성있는 시간대를 부여하고, 아울러 정자직접주입술 (ICSI)을 보다 용이하게 하여 성공적인 수정률과 임신율을 얻음에 있다. 또한 ICSI를 시행한 후, 운동성이 향상된 잉여의 정소정자를 냉동보존함으로써 차후에 TESE을 다시 시행치않고도 시험관 아기 시술을 시도할 수 있는 부가적인 잇점도 있다고 할 수 있다. 대상환자군은 정관폐색증(n=11) 혹은 비정관폐색증(n=2)을 보이는 13명의 무정자증의 남성불임환자였으며 난자회수예정일 3일전에 TESE를 시행하여 정소정자를 얻은 후 이를 정자직접주입술이 시행되는 당일까지 Vero cell과 공배양을 실시하였다. Vero cell과의 공배양에 의하여 운동성이 있는 정소정자의 수는 공배양전과 비교하여 평균 3.3배가 증가하였으며, 특히 공배양전에 운동성이 있는 정소정자의 수가 50,000/ml이하의 미약한 운동성만을 보였던 경우 (n=5)에는 공배양 후에 운동성이 있는 정소정자 수의 평균증가율이 7.7배였다. 공배양전 정자운동성이 전혀 없었던 2례의 비정관폐색증환자중 3일간의 공배양을 통하여 1례에서 운동성을 획득한 정소정자를 얻을 수 있었으며 (14,300/ml), 정자직접주입술을 통하여 성공적인 수정 및 임신에 도달할 수 있었다. Vero cell과 공배양을 하고 ICSI했던 결과, 평균 수정률은 75.0% 이었으며 임신율은 61.5%였다.

• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.307-312
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• 2004

The bioreactor system for the large-scale plant tissue culture was developed to control the pH concentration and DO (dissolved oxygen), and air flowrate. The system controlling the proper air flow rate for each bulblet growth stage and monitoring the contamination of bioreactor using the pH change was controled by computer program. For the uniform bulblet distribution in bioreactor, the proper air flow rate was 300 cc/min at the beginning of bulblet culture, 400 cc/min after 20 days, 500 cc/min after 40 days, 600 cc/min after 60days, and 700 cc/min after 80 days. It was possible to maintain the pH concentration within 5.5$\pm$0.5 during the culture by control system of bioreactor.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.85-85
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• 2001

본 연구는 돼지 난자의 체외 수정시 sucrose 첨가가 돼지 난자의 다정자 침입율에 미치는 영향을 조사하기 위하여 실시하였다. 돼지 난포란은 0.1% PVA, 3.05 mM D-glucose, 0.91 mM sodium pyruvate, 0.57 mM cysteine, 0.5 $\mu\textrm{g}$/$m\ell$ FSH, 0.5 $\mu\textrm{g}$/$m\ell$ LH, 및 10 ng/$m\ell$ EGF가 첨가된 TCM199 배양액에서 42-44 시간 동안 배양하여 체외 성숙을 유도하였으며, 2 mM caffeine과 0.1% BSA가 첨가된 체외수정용 mTBM 배양액에 3% sucorse를 첨가하여 6시간동안 수정을 유도하였다. 수정된 난자의 다정자 침입율을 조사하기 위해 체외수정 후 12, 13 및 14 시간대에 각각 1% orcein으로 염색하여 전핵 형성율을 조사하였다. 돼지 난자의 체외수정시 3% sucorse를 첨가하였을 때, 전핵 형성율을 조사한 결과 대조구에서 미수정, 1, 2, 및 3 PN 이상의 전핵 형성율은 각각 57.6 7.6, 9.7, 및 25%로, 처리구 74.2, 7.6, 13.6, 및 4.5% 에 비교하여 다정자 침입율이 유의하게 높았다(p<0.05). 그리고 3% sucorse를 첨가한 체외수정용 배양액에서 수정을 유도한 후 12, 13, 및 14 시간대별로 나누어 전핵 형성율을 조사한 결과, 수정 후 시간이 경과되면서 다정자 침입율이 증가되는 것을 볼 수 있었다 그러나 대조구 60.7%에 비해서는 각각 32.5, 36.0, 및 33.3%로 현저히 낮은 것을 볼 수 있었다.(Table Omitted). 또한, 3% sucorse가 첨가된 배양액에서 수정된 수정란의 발달율을 조사한 결과, 배반포 생산율은 처리구 21.8%로 대조구 6.9%에 비해 유의하게 높은 것을 확인 하였다. 따라서, 돼지 난자의 체외수정시 3% sucorse첨가는 체외수정란 생산에서 문제가 되고 있는 다정자 침입율을 저하시키고 배발달율을 향상시킴으로서 수정란의 체외 대량생산 효율성은 물론 수정란 이식시 수태율을 증가시킬 수 있을 것으로 사료된다.

• Korean Journal of Medicinal Crop Science
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• v.13 no.4
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• pp.133-137
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• 2005

Experiments were conducted to find the optimal acclimatization conditions for Eleutherococcus senticosus plantlets regenerated from bioreactor cultured somatic embryos, various acclimatizing conditions were compared regarding both survival rate and growth of the plantlets. Among the various temperature and artificial soil tested, the highest survival rate (88%) was observed when plantlets were acclimatized in Klasmann bed soil at $10^{\circ}C$ When in vitro plantlets directly transplanted to field environment, shading treatment was necessary and 50% shading was more effective than 30% shading. Transplanting season were also important for successful acclimatization of in vitro cultured plantlets, transplanted on March 20 with 50% shading exhibited the best survival rate and further growth.

• KSBB Journal
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• v.18 no.2
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• pp.133-139
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• 2003

Established cell-line cultures of cultured and wild mountain ginseng were characterized and their abilities to produce ginsenoside were determined. Cell lines were made of calli induced from the roots of wild mountain ginseng and cultured ginseng(Panax ginseng). Suspension cultures of wild mountain ginseng and cultured ginseng showed different growth and ginsenoside production rate. Their specific growth rates were 0.067 and 0.0035 day-1 in spite of having the same sugar consumption rates, where cells from wild mountain ginseng grew almost twice as fast as those of cultured ginseng. Their respective abilities to produce ginsenoside, however, were 0.53 and 2.53 mg/L.day, which means cells from cultured ginseng produced around 5 times more than wild mountain ginseng.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.63-67
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• 2002

Bifidobacterium strain was isolated from the feces of brast fed infants. The isolated strain was identified as Bifidobacterium longum by 16S rRNA sequence analysis and named as Bifidobacterium SH2. The MRS medium was modified to obtain high density cells of Bifidobacterium SH2. The optimal medium was determined to be 50 g/L lactose, 10 g/L beef extract, 10 g/L peptone, 5 g/L yeast extract, 7 g/L sodium acetate, 2 g/L ammonium citrate, 2 g/L disodium phosphate,1 g/L tween 80, 0.2 g/L MnSO$_4$ and 0.5 g/L L-cysteine. The pH and temperature were optimized as 5.0 and $37^{\circ}C$, respectively. Through out the optimization of medium composition and culture conditions, the dry cell weight and viable cell count were 2.5 times and 1.8 times higer than those in MRS medium, respectively.

• Korean Journal of Plant Tissue Culture
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• v.22 no.4
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• pp.229-234
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• 1995

Anthers of hybrids and inbred lines of Chinese cabbage (Brassica campestris ssp. pekinensis) were inoculated on the modified solid or liquid B$_{5}$ medium after several pre-treatments. Anthers were then maintained at 25 $^{\circ}C$ after being subjected to various post-treatment. Somatic embryos began to appear 9 days after inoculation and ended at 13th days. Low temperature pre-treatment did not increase embryoid production whereas high post-treatment temperature of 4$0^{\circ}C$ or 45$^{\circ}C$ enhance the production. There were considerable levels differences in embryogenesis between lines used, but not between the culture methods. Somatic embryo yield was also increased by subjecting anthers to one day at 35$^{\circ}C$ and then another one day at 3$0^{\circ}C$ after plating. Histological observations showed several stages in haploid development ranging from a few celled to large multiple-celled embryoids.s.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.159-163
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• 1998

In vitro shoot tip culture technique was established in pear (Pyrus pyrifolia 'Niitaka') as related to tree vigor, sampling time, and plant growth regulators and sucrose supplemented to medium. Shoot tips excised in June from the tree having medium-vigor developed good shoots. BA (1.0 and 2.0 mg/L) without NAA produced shoots suitable for proliferation, and NAA supplemented to medium resulted in poor shoot growth and excessive callus formation. BA of 2.0 mg/L combined with 0.01 mg/L NAA provided shoots suitable for rooting and sucrose of 30 g/L was recommended for proliferation medium. A fourth strength MS medium supplemented with 0.1 mg/L NAA produced plantlets in good quality of root number and root length.

• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.116-122
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• 2014

Total of fifty accessions of Brassica rapa with various morphological characteristics were used for production of double haploid plants though microspore culture in Brassica rapa. Among them, only 30 accessions induced embryos from microspores. The highest efficiency of embryo induction of 1.194 per bud was obtained from IT135449 of turnip type, while 3 accessions of sarson (winter oil) type did not generate embryo. The effect of heat shock periods for embryogenesis was also investigated with 4 accessions (IT135449; Turnip type, IT199710; Chinese cabbage type, IT212886; Pak choi type, IT218043; Summer oil type). The high productions of embryos were observed in IT135449, IT199710 and IT212886 when microspores were pre-cultured to $32^{\circ}C$ for 2 days. In IT218043, high embryogenesis was observed at the 3 days of heat shock treatment. The optimal condition of shoot regeneration for IT199710 was observed in MS medium supplemented with NAA $0.5mg{\cdot}L^{-1}$ and BAP $1mg{\cdot}L^{-1}$. In contrast, the IT135449 and IT212886 were observed high regeneration frequency in MS medium without plant growth regulators. All the plantlets regenerated from microspore-derived embryos have been successfully transplanted to soil, and bud self-pollinated seeds were produced from doubled haploid plants. This indicated that double-haploid genotype was likely generated naturally during embryogenesis process.