• Journal of The Korean Society of Grassland and Forage Science
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• v.5 no.2
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• pp.127-135
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• 1985

The effects of morphological development and environmental temperature on synthesis and accumulation behavior of cell-wall constituents were studied in maize cv. Blizzard and sorghum cv. Sioux and Pioneer 931 at Muenchen Technical University from 1979 to 1981. Various growth stages of maize and sorghum plants were grown on field and phytotron at 4 temperature regimes of 30/25, 25/20, 28/18 and 18/8 degree C and mid-summer sunlight over 13-hour days. The results are summarized as follow: 1. Cell-wall constituents in sorghum and maize plants were shown to have a great synthesis rates at early growth stage from growing point differentiation to final leaf visible. The highest concentration of cell wall contents were found at heading stage with 52-54% and 64-68% of neutral detergence fiber, and 30% and 45% of acid detergence fiber foe maize and sorghum, respectively. 2. The structural carbohydrates, cellulose and hemicellulose, were found as a main components of cell-wall constituents. Cellulose were mainly accumulated in stalks, while hemicellulose were an important cell wall components in leaves and panicle. 3. Synthesis rates of cell-wall constituents and non-strnctural carbohydrates were associated with increasing of temperature. Reserved carbohydrates such as fructosan, mono - and dissaccharose in plant were, however, declined when the temperature exceeded 30 deg C, during the accumulation of cellulose, hemicellulose and lignin were increased continuously. 4. Cell-wall constituents lowered digestibility and net energy accumulation in sorghum and maize plants. In a in vitro and in vivo trial, it was found a negative correlation between digestion dry matter and cell wall constituents, especially cellulose and lignin.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.281-287
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• 2013

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

• Korean Journal of Medicinal Crop Science
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• v.8 no.1
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• pp.41-48
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• 2000

This study was aimed to proliferate Gentiana axillariflora Leveille which is one of the important medicinal and ornamental plants, by establishment of multiple shoot formation and embryogenesis through tissue culture technique. Callus was formed on MS (Murashige and Skoog) medium supplemented with 2,4-D, CPA, but not formed with BAP. The addition of 2,4-D 2 mg/ l into the medium was effective for callus formation and the rate of callus formation was about 90%. Somatic embryos were obtained on MS medium for two months. When callus was cultured on MS medium with combination treatment of 2,4-D 0.5 mg/ l and BAP 0.5 mg/ l, the number of embryo formed was better than that of other single or combination treatments and the total numbers of embryo a were 18.8 (number of total embryo/number of explants incubated = 753/40) at mean. Callus induction from stem and node explants was increased by addition of TDZ 2 mg/ l in the presence of 2,4-D 2 mg/ l, respectively. The best result about the differentiation of shoots was obtained on MS medium added BAP 2 mg/ l from node culture. Multiple shoots from shoot apex were induced on MS medium containing BAP 1 mg/ I and TDZ 1 mg/ l , BAP 2 mg/ l and TDZ 1 mg/ l. The number of multiple shoots per one explant was above seventy plants. It was the most effective regeneration system for rapid multiplication of Gentiana axillariflora Leveille.

• KSBB Journal
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• v.20 no.4
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• pp.305-310
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• 2005

Reconstruction of rabbit corneal epithelium was performed through dynamic culture method using self-manufactured amniotic membrane supporter and lyophilized amniotic membrane. Rabbit corneal epithelial cells were cultured and cryopreserved after isolation from limbus, and the cells could be proliferated by passage number 10. The basal layer was well formed, and the epithelium layer was constructed tightly by the increase of cell proliferation and differentiation by dynamic culture method than static culture. Thus, the reconstruction of the corneal epithelium using lyophilized amniotic membrane is considered to be a good in vitro model for transplantation of corneal epithelium to patients with a severely damaged cornea.

• Journal of Periodontal and Implant Science
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• v.39 no.2
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• pp.119-128
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• 2009

Purpose: The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow. Methods: Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation. Results: PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite /${\beta}$- tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation. Conclusions: This study demonstrated that canine dental stem cells have stemness like bone marrow stem cells. Transplantation of these cells might be used as a therapeutic approach for dental stem cell-mediated periodontal tissue regeneration.

• Development and Reproduction
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• v.12 no.1
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• pp.1-8
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• 2008

Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

• Journal of Gastric Cancer
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• v.5 no.3 s.19
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• pp.191-199
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• 2005

Background: Though infections of Helicobacter pylori (H. pylori) are closely associated with activation of host angiogenesis, the underlying mechanisms, as well as the strategy for its prevention, have not been identified. Here, we investigated a causal role of H. pylori infection in angiogenesis of gastric mucosa and a potent inhibitory effect of a gastric proton pump inhibitor (PPI) on the gastropathy. Materials and Methods: A comparative analysis of CD 34 expression in tissues obtained from 20 H. pylori-associated gastritis and 18 H. pylori-negative gastritis patients was performed. Expression of $HIF-1{\alpha}$ and VEGF were tested by using RT-PCR. To evaluate the direct effect of H. pylori infection on differentiation of endothelial HUVEC cells, we carried out an in vitro angiogenesis assay. Results: H. pyfori-associated gastritis tissues showed significantly higher density of $CD34^+$ blood vessels than did H. pylori-negative gastritis tissues, and the levels were well correlated with expressions of $HIF-1{\alpha}$. Conditioned media from H. pylori-infected gastric mucosal cells stimulated a tubular formation of HUVEC cells. We also found a significant inhibitory effect of PPI, an agent frequently used for H. pylori eradication, on H. pylori-induced angiogenesis. This drug effectively inhibited the phosphorylation of MAP kinase ERK1/2, which is a principal signal for H. pylori-induced angiogenesis. Conclusion: The fact that PPls can down-regulate H. pylori-induced angiogenesis suggest that anti-angiogenic treatment using PPI may be a preventive approach for H. pylori-associated carcinogenesis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.6
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• pp.1467-1476
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• 2006

This study was to evaluate the effect of Bulhwangeumjeonggi-san (BS) on mouse Th1 and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of BS extract were measured as well as the amount ${\beta}$-hexosaminidase in RBL-wH3 cells and the levels of TNF-${\alpha}$ and IL-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with BS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total lgE, OVA-specific lgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4- T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of BS extract, it increased proliferation of CD4 cells by 14% in 50 ${\mu}g/m{\ell}$ concentration but it showed an inhibition in higher concentrations. CD4 T cells under Th1/Th2 polarizing conditions for 3 days with BS resulted in mild decrease of IFN- g in Th1 cells and mild increase of IL-4 in Th2 cell at 50 ${\mu}g/m{\ell}$ but the level of IL-4 decreased by 18% at 100 ${\mu}g/m{\ell}$. BS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}$-hexosaminidase in RBL-2H3 cells. Treatment of BS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-${\alpha}$ production. Oral administration of BS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum lgE and OVA-specific lgE by 50% and 55%, respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 25% and significant decrease of IL-4 and IL-5 by 53%, and 38%, respectively. The results show that BS does not strongly induce mouse T cells to transform into Th1 or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.844-852
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• 2006

This study was to evaluate the effect of Yeongyupaedog-san (YGPDS) on mouse Thl and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of YGPDS extract were measured as well as the amount of ${\beta}-hexosaminidase$ in RBL-2H3 cells and the levels of $TNF-{\alpha}$ and 1L-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with YGPDS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total IgE, OVA-specific IgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4 T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of YGPDS extract, it increased proliferation of CD4 cells by 11% in $100\;{\mu}g/^{ml}$ concentration but it showed an inhibition by 37% at $200\;{\mu}g/^{ml}$ CD4 T cells under Th1/Th2 polarizing conditions for 3 days with YGPDS resulted in mild decrease of IFN- g in Thl cells and significant decrease of IL-4 in Th2 cells at $500\;{\mu}g/^{ml}\;and\;100\;{\mu}g/^{ml}$ by 18% and 21%, respectively. YGPDS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}-hexosaminidase$ in RBL-2H3 cells. Treatment of YGPDS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-n production. Oral administration of YGPDS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum IgE and OVA-specific IgE by 25% and 34% , respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 44% and significant decrease of IL-4 and IL-5 by 56%, and 24%, respectively. The results show that YGPDS does not strongly induce mouse T cells to transform into Thl or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

• Journal of the East Asian Society of Dietary Life
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• v.25 no.1
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• pp.99-110
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• 2015

The purpose of this study was to evaluate the inhibitory effects of commonly consumed vegetables in Korea on lipid accumulation and production of pro-inflammatory cytokines related to obesity/metabolic syndrome. Using KNHANES (Korea National Health and Nutrition Examination Survey) raw data ($1^{st}$; 1998, $5^{th}$; 2010, 2011) and a literature search, we selected vegetables for study. Edible portions of samples were prepared, ethanol-extracted, and then freeze-dried. 3T3-L1 adipocytes and HepG2 hepatocytes cells were used as in vitro models. Lipid accumulation determined by Oil-red O staining showed that all samples except bracken had inhibitory effects on lipid accumulation in 3T3-L1 adipocytes. Especially, crown daisy and mugwort effectively reduced accumulation of lipids, and their inhibition rates were more than 60% of the control group. Young pumpkin, leeks, crown daisy, and mugwort showed significantly decreased MCP-1 levels compared to the control group. However, adiponectin protein level did not increase in the vegetables experimental group. In HepG2 hepatocytes, all samples showed inhibitory effects on lipid accumulation at one of the two concentrations. Although adiponectin protein levels did not increase, MCP-1 protein levels decreased in adipocytes. Further, lipid accumulation in adipocytes and hepatocytes decreased. In conclusion, all samples showed one or more improved obesity/metabolic syndrome indicators. Among them, young pumpkin, leeks, crown daisy, and mugwort were selected as the most effective portions of vegetables based on improvement of obesity/metabolic syndrome-related indicators.