• Analytical Science and Technology
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• v.27 no.1
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• pp.27-33
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• 2014

This study described about preparation of the cross-linked silk fibroin (SF) film by ${\gamma}$-irradiation of the casted SF film, which is fabricated from aqueous solution regenerated via fibers of cocoons and their application as supports for human cell culture. The properties of cross-linked SF film were evaluated by FT-IR spectroscopy, contact angle, solubility to water, thermal analysis, surface area analyzer, and morphology via scanning electron microscopy (SEM). The cross-linked SF films were not dissolved in water and exhibited the rough surface morphology, large surface area, and good thermal properties. The human fibroblast cell (CCD-986sk) and embryo kidney-ft cell were well growed on the surface of cross-linked SF film supports prepared by ${\gamma}$-irradiation. The cross-linked SF film prepared by ${\gamma}$-irradiation can be used as biomaterials for human cell culture.

• Journal of Pharmaceutical Investigation
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• v.40 no.6
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• pp.321-332
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• 2010

Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the "passage" culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.

• Development and Reproduction
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• v.16 no.4
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• pp.353-361
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• 2012

Human embryonic stem (ES) cells are a potential source of cells for developmental studies and for a variety of applications in transplantation therapies and drug discovery. However, human ES cells are difficult to culture and maintain at a large scale, which is one of the most serious obstacles in human ES cell research. Culture of human ES cells on MEF cells after disassociation with accutase has previously been demonstrated by other research groups. Here, we confirmed that human ES cells (H9) can maintain stem cell properties when the cells are passaged as single cells under a feeder-free culture condition. Accutase-dissociated human ES cells showed normal karyotype, stem cell marker expression, and morphology. We prepared frozen stocks during the culture period, thawed two of the human ES cell stocks, and analyzed the cells after culture with the same method. Although the cells revealed normal expression of stem cell marker genes, they had abnormal karyotypes. Therefore, we suggest that accutase-dissociated single cells can be usefully expanded in a feeder-free condition but chromosomal modification should be considered in the culture after freeze-thawing.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.337.3-338
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• 2002

UV-B irradiation increases the synthesis of matrixmetalloproteinase-1 (MMP-1) that degrades skin collagen in human skin. In this work, we investigated the photoprotective effect of decursinol angelate (DEA) extracted Irom Angelica gigas on human skin libroblasts. DEA inhibited UVB-induced MMP-1 induction, which was conlirmed by western blot and ELISA. We examined upstream signal transduction pathway and the action mechanism of DEA on UVB induction of MMp in human skin fibroblasts. (omitted)

• Proceedings of the Botanical Society of Korea Conference
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• 1985.08b
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• pp.23-33
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• 1985

Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

• Proceedings of the Botanical Society of Korea Conference
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• 1999.07a
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• pp.45-49
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• 1999

Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.140.1-140.1
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• 2003

Permanent cell culture lines derived from human cancer tissue are important experimental models in the study of human cancer cell proliferation. The in vitro effects of C. militaris and its extracted fractions on the human breast cancer (SK-BR3), liver cancer (SK-Hep1, HepG2), kidney cancer (p15), lymphoma (Jurkat) were studied. F1 (CCCA, crude cordycepin containing adenosine), F2 (ethanol precipitation), F3 (ethanol soluble supernatant) and F4 (fraction of through SK-1B) significantly stimulated in vitro cytotoxic in human cancer cell lines. (omitted)

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.25-40
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• 2006

Purpose : This study was undertaken to evaluate the inhibitory effects of Arisaematis rhizoma on the cell proliferation in HeLa cells. Methods : The cultured cell after treatment in the different duration in 24, 48, 72 hours with solution of 1%. 5%, 10% Arisaematis rhizoma was quantified by trypan blue exclusin method. The control group was treated with 2% FBS in the different duration in 24, 48, 72 hours. We examined DNA of activated caspase by FACS analysis, caspase-3 activity, DNA fragmentation by DNA laddering, activity of HeLa Cells by the XTT assay, activity of MAP kinase by RT-PCR analysis. Results : After 72 hours culture, the growth activities of 1%, 5%, 10% Arisaematis rhizoma-treated Hela cell were significantly reduced with control group, respectively. After 24 hours culture, the ratio of cells showing caspase activity by FACS analysis were increased in 1%, 5%, 10% Arisaematis rhizoma-treated Hela cell. It were also increased in 48 hours culture of 10% and 72 hours culture of 5%, 10% Arisaematis rhizoma-treated Hela cell. In 24, 48 and 72 hours culture, DNA fragmentations of 5%, 10% Arisaematis rhizoma-treated Hela cell were obviously observed. These results meaned that Arisaematis rhizoma induces apoptosis of HeLa cells. It was supported by increased caspase-3 activity and decreased MAP kinase activity according to time periods and concentrations of Arisaematis rhizoma solution. Conclusion : The study shows that Arisaematis rhizoma has inhibitory effect on cell proliferation and induction capacity of apoptosis of human cevical carcinoma cell line, HeLa cells, in vitro. These results suggest that Arisaematis rhizoma should be useful for treatment of human cevical carcinoma.

• Archives of Plastic Surgery
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• v.32 no.3
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• pp.343-346
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• 2005

It has been established that a graft of fibroblasts is able to improve wound healing. However, there has been no research on the effect of a graft of bone marrow stromal cells on wound healing. The wound healing process requires cell proliferation and production of extracellular matrix and various growth factors. The purpose of this study was to compare the abilities of human fibroblasts and bone marrow stromal cells, which contains mesenchymal stem cells, to proliferate and to produce collagen. Human bone marrow stromal cells and fibroblasts were isolated from bone marrow and dermis of the same patients and grown in culture respectively. Cell proliferation and production of type I collagen by human bone marrow stromal cells and dermal fibroblasts were examined by MTT method and by ELISA of cell culture media on day 1, 3, and 5 days post-incubating. The human bone marrow stromal cells showed 11-17% higher cell proliferation than fibroblasts at each time interval. The levels of type I collagen in the human bone marrow stromal cell group was also significantly higher than those in the fibroblast group. The results indicate that the grafts of human bone marrow stromal cells can show more promising effect than that of fibroblasts for healing of chronic wounds.

• Reproductive and Developmental Biology
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• v.28 no.4
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• pp.247-252
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• 2004

Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.