• Journal of Plant Biotechnology
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• 제32권3호
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• pp.209-215
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• 2005

Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

• Journal of Life Science
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• 제20권12호
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• pp.1820-1828
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• 2010

Cobalt(II) chloride, a chemical compound with the formula$CoCl_2$, has been widely used in the treatment of anemia, as a chemical agent for the induction of hypoxia in cell cultures, and is known to activate hypoxic signaling. However, excessive exposure to cobalt is associated with several clinical conditions, including asthma, pneumonia, and hematological abnormalities, and can lead to tissue and cellular toxicity. It is also known to induce apoptosis. One of the questions was that of whether $CoCl_2$ might induce apoptosis via endoplasmic reticulum (ER) stress in neurons. To address this question, first, the level of DNA fragmentation was measured for assay of apoptotic rates using $CoCl_2$ with neuron PC12 cells. After confirmation of apoptosis inductions, under the same conditions, the expression levels of ER stress associated factors [ER chaperones Bip, calnexin, ERp72, ERp29, PDI, and ER membrane kinases (IRE1, ATF6, PERK)] were examined by RT-PCR and Western blotting. These results indicated that apoptosis is induced through activation of ER membrane kinases via ER stress. In conclusion, during induction of apoptosis through $CoCl_2$-induced hypoxia in neuron PC12 cells, ER membrane kinase of IRE1 was dominantly up-expressed, and, consecutively, TRAF2, which has been suggested to be one of the links connecting apoptosis and ER stress, was strongly up-expressed.

• Clinical and Experimental Reproductive Medicine
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• 제43권3호
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• pp.146-151
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• 2016

Objective: This study was conducted to compare the effects of static culture, dynamic culture, and the combination of dynamic culture with specialized surfaces involving co-culture on human embryonic development. Embryos cultured using conventional static culture (SC) techniques served as a control group. We compared dynamic culture using micro-vibration culture (MVC) and micro-vibration with co-culture (MCoC), in which autologous cumulus cells were used as a specialized surface. Methods: We conducted a chart review of patients who were treated between January 2011 and November 2014 in order to compare embryonic development rates and pregnancy rates among the groups. Zygotes were cultured in micro-droplets, and embryos were subsequently selected for transfer. Some surplus embryos were cryopreserved, and the others were cultured for blastocyst development. A micro-vibrator was set at the frequency of 42 Hz for duration of 5 seconds per 60 minutes to facilitate embryo development. Results: No significant differences among the groups were present in patient's characteristics. However, the clinical pregnancy rates were significantly higher in the MVC group and the MCoC group than in the SC group. No significant differences were found in the blastocyst development rate between the SC group and the MVC group, but the blastocyst development rate in the MCoC group was significantly higher than in the SC and MVC groups. Conclusion: The clinical pregnancy rate was significantly increased by the application of micro-vibration to the embryonic cultures of poor responders. The blastocyst development rate was significantly increased by the application of MCoC to surplus embryos.

• YAKHAK HOEJI
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• 제49권6호
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• pp.459-464
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• 2005

The immune reinforcement of the probiotic lactic acid bacteria Lactobacillus acidophilus, Streptococcus thermophilus and Bifidobacterium bifidum was studied in RAW 264.7 cell line treated with diluted solution (dilution to $2^{5}$) of the supernatnats of lactic acid bacteria. RAW 264.7 cell line was used as a macrophage model to assess the effects of lactic acid bacteria on the production of nitric oxide (NO) and cytokine tumor necrosis factor (TNF)-$\alpha$ and cell morphological changes. The production of NO and TNF-$\alpha$ were largely affected by lactic acid bacteria in dose-dependent manner in 24 or 48 hr cultures and cell morphological changes were also largely affected by lactic acid bacteria. Especially Bifidobacterium bifidum differentially stimulated the production of NO and TNF-$\alpha$. NO production was increased by Bifidobacterium bifidum 25 $\mu$l/ml more than LPS (20 ng/ml) control, and TW-$\alpha$ by Bifidobacterium bifidum 6.25 $\mu$l/ml more than LPS (10 ng/ml) control. The in vitro approaches employed here should be useful in further characterization of the effects of lactic acid bacteria on systemic immunity.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 한국정보통신학회 2018년도 추계학술대회
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• pp.448-451
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• 2018

The purpose of this study was to investigate the developmental process of myelination by neuron and Schwann cell cultures and the development of demyelination by herpes simplex virus-1 infection by electron microscopy and molecular biological analysis. The dorsal root ganglion (DRG) was isolated from the mouse embryo and Schwann cells and neuronal cells were cultured in vitro. Neuronal cells treated with mitotic inhibitors and purified Schwann cells were co-cultured together to induce myelination. The herpes simplex virus-1 was infected with the co-cultured cells, and the demyelination was induced. The myelin protein zero (MPZ) antibody, which means the presence of myelin formation, was used and electron microscopy was used to observe the development of myelin and dehydration.

• KSBB Journal
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• 제13권2호
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• pp.187-194
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• 1998

For mass production of polyhydroxyalkanoates (PHA), high cell density cultures of Alcaligenes eutrophus by fed-batch culture under phosphate-limitation condition has been investigated. PHA accumulation by the regulation by the regulation of initial phosphate concentration could be automatically induced, and high density cell culture above 200 g/L also could be successfully produced. The production of Poly-$\beta$-hydroxybutyrate (PHB) and dry cell weight increased with increasing the initial phosphate concentration. When the initial concentrations of phosphate were in the ranges of 1.5~4.5 g-PO$_4$/L, PHB and dry cell weight obtained were 83~266 g/L and 61~216 g/L, respectively, and PHB productivity was in the ranges of 1.35~3.10 g/L.h. When a mixture of glucose and propionic acid is used as carbon sources, poly(3-hydroxybutyrate-co-poly-3-hydroxyvalerate), P(3HB-co-3HV), could be also successfully produced under phosphate limitation condition. When the mole ratio of propionic acid to glucose in the feeding solution is 0.22, a final dry cell weight of 150 g/L and a P(3-HB-co-3HV) of 90 g/L were produced. Morphological changes and size distribution of PHB granules synthesized in A. eutrophus under phosphate-limitation condition are determined by TEM during the course of fed-batch. Mean granule diameters of PHB produced are in the range of 0.36~0.39 $\mu$m, and mean cell size was elongated from 0.54~0.59 $\mu$m$\times$ 1.3~1.5 $\mu$m to 0.83~0.89 $\mu$m $\times$2.0~2.3 $\mu$m. Phosphate concentration in media did not affect size distribution of PHB granule and cell.

• Korean Journal of Plant Resources
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• 제27권3호
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• pp.242-248
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• 2014

This study was conducted to establish an efficient transformation system by using somatic embryogenesis in an important Korean native conifer, Korean fir (Abies koreana). Embryogenic masses were induced from mature zygotic embryos of the Korean fir on Schenk and Hildebrandt medium, which was supplemented with thidiazuron. For genetic transformation, the embryogenic masses were co-cultivated with a disarmed Agrobacterium tumefaciens strain C58/pMP90 containing the plasmid vector pBIV10 or LBA4404 containing the plasmid vector MP90. Both vectors contain the kanamycin resistance and beta-glucuronidase (GUS) reporter genes. A total of 48 lines of embryogenic masses were selected on mLV medium containing $50{\mu}g/mL$ of kanamycin after 4 weeks of culture, following 3 days of co-cultivation with A. tumefaciens strain C58/pMP90 carrying pBIV10 (none of the lines was cultivated with strain LBA4404 carrying MP90). Quantitative real-time PCR was performed, and high levels of GUS transcripts were observed in the 48 putative transgenic lines; however, the control (non-transgenic line) showed negative results. Results of histochemical staining showed that the expression of the GUS reporter gene was observed in somatic embryos that developed from the embryogenic masses of all 48 lines. Stably transformed cultures were successfully produced by co-cultivation with A. tumefaciens strain C58/pMP90 carrying pBIV10 in Korean fir. Here, we have reported an Agrobacterium-mediated gene transfer protocol via somatic embryogenesis that may be helpful in developing breeding and conservation strategies for the Korean fir.

• Journal of Microbiology and Biotechnology
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• 제32권3호
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• pp.378-386
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• 2022

Scenedesmus obliquus ABC-009 is a microalgal strain that accumulates large amounts of lutein, particularly when subjected to growth-limiting conditions. Here, the performance of this strain was evaluated for the simultaneous production of lutein and biofuels under three different modes of cultivation - photoautotrophic mode using BG-11 medium with air or 2% CO₂ and heterotrophic mode using YM medium. While it was found that the highest fatty acid methyl ester (FAME) level and lutein content per biomass (%) were achieved in BG-11 medium with CO₂ and air, respectively, heterotrophic cultivation resulted in much higher biomass productivity. While the cell concentrations of the cultures grown under BG-11 and CO₂ were largely similar to those grown in YM medium, the disparity in the biomass yield was largely attributed to the larger cell volume in heterotrophically cultivated cells. Post-cultivation light treatment was found to further enhance the biomass productivity in all three cases and lutein content in heterotrophic conditions. Consequently, the maximum biomass (757.14 ± 20.20 mg/l/d), FAME (92.78 ± 0.08 mg/l/d), and lutein (1.006 ± 0.23 mg/l/d) productivities were obtained under heterotrophic cultivation. Next, large-scale lutein production using microalgae was demonstrated using a 1-ton open raceway pond cultivation system and a low-cost fertilizer (Eco-Sol). The overall biomass yields were similar in both media, while slightly higher lutein content was obtained using the fertilizer owing to the higher nitrogen content.

• International Journal of Stem Cells
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• 제16권2호
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• pp.244-249
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• 2023

Background and Objectives: To examine whether ischemic retinal ganglion cells (RGCs) will be salvaged from cell death by human adipose-derived mesenchymal stem cells (ADSCs) in an organotypic retina model. Methods and Results: Deprived of arterial oxygen supply, whole mice retinas were cultured as an ex vivo organotypic cultures on an insert membrane in a 24-well plate. The therapeutic potential of ADSCs was examined by co-culture with organotypic retinas. ADSCs were seeded on top of the RGCs allowing direct contact, or at the bottom of the well, sharing the same culture media and allowing a paracrine activity. The number of surviving RGCs was assessed using Brn3a staining and confocal microscopy. Cytokine secretion of ADSCs to medium was analyzed by cytokine array. When co-cultured with ADSCs, the number of surviving RGCs was similarly significantly higher in both treatment groups compared to controls. Analysis of ADSCs cytokines secretion profile, showed secretion of anti-apoptotic and pro-proliferative cytokines (threshold>1.4). Transplantation of ADSCs in a co-culture system with organotypic ischemic retinas resulted in RGCs recovery. Since there was no advantage to direct contact of ADSCs with RGCs, the beneficial effect seen may be related to paracrine activity of ADSCs. Conclusions: These data correlated with secretion profile of ADSCs' anti-apoptotic and pro-proliferative cytokines.

• Journal of Plant Biotechnology
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• 제34권1호
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• pp.1-6
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• 2007

Effect of BA concentrations and culture methods on in vitro plant multiplication from shoot-tip cultures of Wasabia japonica was studied. Shoot-tips with leaf primordia and apical meristem were cultured on MS basal medium for all the experiments. Liquid medium for 2 weeks followed by semi-solid medium for 4 weeks containing 1.0 mg/L BA was the best to number of shoots (22.8) and shoot length (3.5 cm). Shoots proliferated could be divided into ca. 5 to 11 of cultures for the multiplication of plantlets. Divided plantlets showed root formation (90%) well onto MS basal medium without growth regulators like IBA and NAA. After rooting, all the plantlets transferred into the pots containing composed soil (bio-media Co., peatmoss $8{\sim}10%$, coir dust $66{\sim}70%$, zeolite $13{\sim}17%$, vermiculite $3{\sim}7%$, perlite $2{\sim}4%$) and grown well into whole plants with multiple shoots.