• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.60-60
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• 2003

Embryonic stem (ES) cells have property of self-renewal and can differentiate into the cells of all three primary germ layers. Recently, many growth factors, alteration of culture condition and gene modifications have been used to differentiate mouse and human ES cells into specific cell types. This study was performed to evaluate the differentiation protocol for human ES cells to the endodermal lineage cells. Human ES cells (Miz-hESl ) were cultured on STO feeder layer mitotically inactivated with mitemycin C, and embryoid bodies (EBs) were formed by suspension culture. Differentiation protocol of EBs consisted of three steps: stage I, culture of EBs for 6 days with ITSFn medium; stage II, culture of stage I cells for 8 days with N2 medium ; stage III, culture of stage II cells for 22 days with N2 medium. mRNA levels of the endodermal lineage differentiation genes were analyzed by semi- quantitative RT-PCR. The Oct-4 expression, a marker of the pluripotent state, was detected in undifferentiated human ES cells but progressively decreased after EBs formation. Differentiating human ES cells expressed marker genes of endodermal differentiation and pancreatic islet cells. GATA4, a-fetoprotein, Glut-2, and Ngn3 were expressed in all stages. However, albumin and insulin were expressed in only stage III cells. The human ES cells can be differentiated into endodermal lineage cells by multiple step culture system using various supplements. We are developing the more effective protocols for guided differentiation of human ES cells.

• Journal of Embryo Transfer
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• v.27 no.3
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• pp.189-192
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• 2012

Although embryonic stem cells (ESCs) or ES-like cells are reported from many mammalian species other than the mouse, the culture system for murine ESCs may not be suitable to the other species. Previously many other research groups have modified either human or mouse ESC culture systems for bovine ESC culture. In this study, we compared three different culture mediums consisting of DMEM, ${\alpha}$-MEM or KnockOut$^{TM}$-DMEM (KO), which are modified from human or mouse ESC culture system, for the generation of bovine ESCs. In this study, some pre-requisite events which are important for establishment and long-term propagation of ESCs such as inner cell mass (ICM) attachment on feeder cells, primary colony formation and sustainability after passaging. Once the ICM clumps attached on feeder cells, this was designated as passage 0. In regards to the rate of ICM attachment, ${\alpha}$-MEM was superior to the other systems. For primary colony formation, there was no difference between DMEM and ${\alpha}$-MEM whereas KO showed lower formation rate than the other groups. For passaging, the colonies were split into 2~4 pieces and passed every 5~6 days. From passage 1 to passage 3, DMEM system seemed to be appropriate for maintaining putative bovine ESCs. On the other hand, ${\alpha}$-MEM tended to be more suitable after passage 6. Although ${\alpha}$-MEM support to maintain a ES-like cell progenies to passage 15, all three culture systems which are modified from human or mouse ESC culture media failed to retain the propagation and long-term culture of putative bovine ESCs. Our findings imply that more optimized alternative culture system is required for establishing bovine ESC lines.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.126-129
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• 1991

In order to produce ${\gamma}-linolenic$ acid by Mucor sp. KCTC 8405P. the fungus was cultivated in fed-batch culture with two phases. i.e., growth in yeast-like form and induction to hyphal growth by pH shift of the culture medium during cultivation. The synchronous growth of the fungus into the appropriate sizes was important for the high density cell culture of this dimorphic fungus. Dissolved oxygen concentration in the medium did not affect degree of unsaturation of fatty acids and ${\gamma}-linolenic$ acid content. Under the culture conditions applied in this experiment. the fungus was found to produce 100 g/l dry mycelia containing 40% of the lipids, where ${\gamma}-linolenic$ acid comprised about 9% of the total extractable fatty acids.

• Journal of Microbiology and Biotechnology
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• v.24 no.5
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• pp.629-638
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• 2014

This study describes an alternative mixed culture fermentation technology to anaerobically convert lignocellulosic biomass into butyric acid, a valuable product with wide application, without supplementary cellulolytic enzymes. Rice straw was soaked in 1% NaOH solution to increase digestibility. Among the tested pretreatment conditions, soaking rice straw at $50^{\circ}C$ for 72 h removed ~66% of the lignin, but retained ~84% of the cellulose and ~71% of the hemicellulose. By using an undefined cellulose-degrading butyrate-producing microbial community as butyric acid producer in batch fermentation, about 6 g/l of butyric acid was produced from the pretreated rice straw, which accounted for ~76% of the total volatile fatty acids. In the repeated-batch operation, the butyric acid production declined batch by batch, which was most possibly caused by the shift of microbial community structure monitored by denaturing gradient gel electrophoresis. In this study, batch operation was observed to be more suitable for butyric acid production.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.106-109
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• 2000

Vascular endothelial cells (EGs) are usually difficult to culture to culture in a large scale because of their complicated requirements for cell growth. As the vascular endothelial growth factor (VEGF) is a key growth factor in the EC culture, we transfected human umbilical vein endothelial cells (HUVEC) using a plasmid containing VEGF gene and let them grow in a culture medium eliminated an important supplement, endothelail cell growth supplement(ECGS). The expression of VEGF by HUVEC tansfected with Vegf GENE was not enough to stimulate the growth of HUVEC, only 40% of maximum cell density obtainable in the presence of ECGS. However, when the culture medium was supplied with 2.5 ng/ml of basic fibroblast growth factor (bFGF), a synergistic effect effect of VEGE and bFGF was observed. In this case, the final cell density was recovered was recovered up to about 78% of maxium value.

• Journal of Microbiology and Biotechnology
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• v.8 no.5
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• pp.461-465
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• 1998

The suspension culture of an anchorage-dependent cell line (Bok-l) from Bombina orientalis was successful in respects of cost and efficiency. The amount of cells obtained from the suspension culture was almost equivalent to that from the anchorage-dependent culture. This result shows the possibility of suspension culture for scale-up. The cells in suspension produced an antibacterial peptide as much as anchorage-dependent cells did. The cell growth ($6.0\times10^6cells/m\ell$) and viability (>80%) at 10 rpm were higher than that at 0 rpm ($1.9\times10^6cells/m\ell$, 65~80%) and 30 rpm ($1.8\times10^6cells/m\ell$ 40~76%). The size of cells became smaller at the agitation rate of 30 rpm. The antibacterial activities of cell extracts from suspension cultured cells were confirmed against gram-negative and gram-positive bacteria by the inhibition zone assay and the liquid growth inhibition assay.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1314-1321
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• 2006

The objective of this work was to study the effect of mixed culture solid substrate fermentation of C. tropicalis ZD-3 with A. niger ZD-8 on detoxification of cottonseed meal (CSM), and to investigate the effect of fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment on the reduction of free gossypol levels during mixed culture solid substrate fermentation of CSM. Experiment 1: Three groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. tropicalis ZD-3, A. niger ZD-8 or mixed culture (C. tropicalis ZD-3 with A. niger ZD-8). One non-inoculated group was used as the control. Levels of initial and final free gossypol (FG), CP and in vitro CP digestibility were assayed. The results indicated that mixed culture fermentation was far more effective than single strain fermentation, which not only had higher detoxification rate, but also had higher CP content and in vitro digestibility. Experiment 2: CSM substrates were treated according to experimental variables including fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment, Then, the treated CSM substrates were inoculated with mixed culture (C. tropicalis ZD-3 with A. niger ZD-8) and incubated at $30^{\circ}C$ for 36 h in a 95% relative humidity chamber. After fermentation ended, FG and CP content of fermented CSM substrate was assayed. The results showed that the appropriate fermentation period was 36 h, and the optimal proportion of CSM in substrate was 70%. Addition of sodium carbonate to CSM substrate was beneficial for fermentative detoxification. Heat treatment could facilitate fermentative detoxification, and supplementation with minerals was instrumental in reducing gossypol levels during mixed culture solid substrate fermentation of CSM.

• Journal of Forest and Environmental Science
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• v.34 no.5
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• pp.395-404
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• 2018

Since the mass mortality of Quercus mongolica has been first reported in Gyeonggi province at 2004, the disease spread rapidly over Korean peninsula annually. Ambrosia beetle (Platypus koryoensis) was known as the insect vector of oak wilt fungus, Raffaelea quercus-mongolicae, and control methods of the disease had mainly been focused on eradication of insect vector. However, for the efficient management of the disease, combined control methods for both of the pathogenic fungus and insect vector are strongly required. As one of the efforts to suppress the pathogenic fungus, antifungal activities of Streptomyces isolated from oak forest soil were assayed in this study. Optimum culture condition for the selected isolates was also studied, As a result, Streptomyces blastmyceticus cultured in PDB (Potato Dextrose Broth) at $25^{\circ}C$ for 1 week showed the strongest antifungal activity against oak wilt fungus. Mycelial growth inhibition rates (MGIRs) of Streptomyces isolates were compared on culture media supplemented with heated and unheated culture filtrates of S. blastmyceticus. MGIRs on culture media with unheated culture filtrates were generally higher than those on culture media with heated culture filtrates. Antagonistic mechanism to get involved in the inhibition of hyphal growth and spore formation of the pathogen is due to the antifungal metabolites produced by Streptomyces. This study will provide the fundamental information in developing biocontrol agents for the environment-friendly management of oak wilt disease.

• Journal of Plant Biotechnology
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• v.8 no.1
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• pp.27-35
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• 2006

Plantlet regeneration in Asteracantha longifolia(L.) Nees (Acanthaceae), a medicinal herb has been achieved from seedling explants on basal MS medium. Three different seedling explants including node, internode and leaf segments on used. Of these three explant, leaf explants gave better response for both callus mediated organogenesis and direct multiple shoot induction. Number of explants showing differentiation of shout buds was higher on MS media supplemented with BA compared to kinetin. MS medium fortified with BA ($2.0mgl^{-1}$) and NAA ($0.5mgl^{-1}$) was found to be most suitable for both callus mediated organogenesis and elongation of shouts. The elongated shoots were successfully routed on MS medium fortified with NAA or IBA. Among them $0.1mgl^{-1}$ NAA or $0.2mgl^{-1}$ IBA provides better response for rhizogenesis. Regenerated plantlets were successfully established in soil where 85.4% or them developed into morphologically normal and fertile plants. RAPD profiling using four decamer primers confirmed the genetic uniformity of the regenerated plantlets and substantiated the efficacy and suitability of this protocol for in vitro propagation of A. longifolia.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.3
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• pp.152-157
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• 2006

The purpose of this study was to evaluate the usefulness of the automated TB-PCR assay for the detection of Mycobacterium tuberculosis. The 807 cases were analyzed with their TB-PCR, AFB smear and culture in bronchial washing fluids, sputum and body fluids samples. The TB-PCR positive of the bronchial washing fluid, sputum and body fluids were 11.3%, 7.3% and 3.6%, respectively, in cases of AFB smear-negative and culture-negative. The sensitivity values of the bronchial washing fluid, sputum and body fluids were 93.3%, 100% and 50%, respectively, according to the culture result. The sensitivity of body fluids was lower than that of the bronchial washing fluid and sputum. The specificity values of the bronchial washing fluid, sputum and body fluids were 83.3%, 89.0% and 95.7%, respectively, according to the culture result. In conclusion, the automated TB-PCR assay proved to be a useful method for the detection of Mycobacterium tuberculosis in the bronchial washing fluid and sputum. But we think that there is still a need for us to study body fluids further.