• Title/Summary/Keyword: stem culture

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Study into prosodic morphological analysis in Korean and Standard Chinese partial reduplication and contrastive analysis using optimality theory (한국어와 표준 중국어 부분 중첩 의성·의태어에 대한 운율 형태론적 분석의 문제와 최적성 이론적 분석을 통한 대조 분석)

  • Chang, Jae-Woong
    • Cross-Cultural Studies
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    • v.49
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    • pp.275-301
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    • 2017
  • Based on the previous analysis by Wanghongjun(2008), this paper applies Optimality Theory (OT) to the field of partial reduplication in Korea to increase its scientific validity. Toward this end, I propose an alternative analysis of Korean partial reduplication, applied as a single process of prefixation. Reduplicated words by prefixation are divided into two types: with a heavy syllable of the stem, and another with the stem. The two types of partial reduplication are closely related by the sound features. In addition, I discussed Chinese partial reduplication from a prefixation perspective within the framework of OT and performed contrastive analysis of Korean and Standard Chinese processes of reduplication by focusing on constrained rankings. As a result, the alternative analysis showed a systematic relationship among the reduplicated words in Korean and Standard Chinese. Lastly, I explained that the coronal /t/ and liquid /l/ have a special functional hierarchy in both languages. This study can be re-explained based on the framework of OT according to a few major constraints, involving MAX-BR, MAX-IO. Their rankings show the adequacy of analysis.

Effect of LPS and melatonin on early development of mouse embryo

  • Park, Haeun;Jang, Hoon;Choi, Youngsok
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.3
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    • pp.183-192
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    • 2022
  • Lipopolysaccharide (LPS) is an endotoxin factor present in the cell wall of Gram-negative bacteria and induces various immune responses to infection. Recent studies have reported that LPS induces cellular stress in various cells including oocytes and embryos. Melatonin (N-acetyl-5-methoxytryptamine) is a regulatory hormone of circadian rhythm and a powerful antioxidant. It has been known that melatonin has an effective function in scavenging oxygen free radicals and has been used as an antioxidant to reduce the cytotoxic effects induced by LPS. However, the effect of melatonin on LPS treated early embryonic development has not yet been confirmed. In this study, we cultured mouse embryos in medium supplemented with LPS or/and melatonin up to the blastocyst stage in vitro and then evaluated the developmental rate. As a result of the LPS-treatment, the rate of blastocyst development was significantly reduced compared to the control group in all the LPS groups. Next, in the melatonin only treated group, there was no statistical difference in embryonic development and no toxic effects were observed. And then we found that the treatment of melatonin improved the rates of compaction and blastocyst development of LPS-treated embryos. In addition, we showed that melatonin treatment decreased ROS levels compared to the LPS only treated group. In conclusion, we demonstrated the protective effect of melatonin on the embryonic developmental rate reduced by LPS. These results suggest a direction to improve reproduction loss that may occur due to LPS exposure and bacterial infection through the using of melatonin during in vitro culture.

Effect of Coir Substrate Mixing Ratios on the Growth and Yield of Perilla Leaves under Hydroponics (수경재배 잎들깨의 생육과 수량에 미치는 코이어 배지의 혼합비율 효과)

  • Pyeong-Sic Park;Jong-Won Park;Hye-Kyeong Hyeon;Hyun-Sook Kim;Soo-Sang Hahm;Hak-Hun Kim;Hyo-Gil Choi
    • Journal of Environmental Science International
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    • v.33 no.1
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    • pp.17-25
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    • 2024
  • This study aimed to determine the most suitable coir substrate mixing ratio for optimizing the growth and yield of the "lpduelkkae 1" cultivar. We comprehensively analyzed the physicochemical properties, growth, and yields of four different substrate combinations: perlite (coir with mixing ratios of 70:30 (PC30), 50:50 (PC50), and 30:70 (PC70)) and 100% coir (C100). The results revealed substantial differences in substrate properties. C100 exhibited the highest total porosity and the lowest solid phase, indicating excellent air permeability. The pH levels and electrical conductivity (EC) values ranged from 5.4-6.8 and 1.2-3.1 dS·m-1, respectively. Leaf growth parameters, including length, width, and dry weight, showed positive correlations with high coir ratios, except for PC30. PC70 and C100 outperformed other substrates in stem growth, exhibiting superior stem diameter and fresh and dry weights. The quantity of marketable leaves was the highest in the C100 substrate. Furthermore, C100 comprised integrated levels of essential nutrients, such as Ca and Mg, owing to its high coir content. In conclusion, a coir ratio of approximately 70% (v/v) should be maintained in the substrate for creating an optimal cultivation environment. Furthermore, the selection of humidity-resistant varieties as well as precise nutrient and moisture management for different seasons and growth stages are crucial for a successful perilla leaf hydroponic cultivation.

Establishing porcine jejunum-derived intestinal organoids to study the function of intestinal epithelium as an alternative for animal testing

  • Bo Ram Lee;Sun A Ock;Mi Ryung Park;Min Gook Lee;Sung June Byun
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.1
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    • pp.2-11
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    • 2024
  • Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

Mouse Embryonic Stem Cell Uptakes of Buforin 2 and pEP-1 Conjugated with EGFP (생쥐 배아 줄기세포의 Buforin 2 및 pEP-1에 결합된 EGFP의 세포 내 수송)

  • Jung, Su-Hyun;Park, Seong-Soon;Lim, Hyun-Jung;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.11 no.2
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    • pp.111-119
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    • 2007
  • Differentiation of cells can be induced through modulation of endogenous regulators using exogenous factors. Useful transfection systems to transport a specific exogenous regulator into cell have been tried but still there are many obstacles to overcome. In this study, we examined the transfection efficiency of cell permeable peptides (CPPs) in mouse embryonic stem cell under the various conditions. To identify the CPP-mediated translocation of a protein, we employed recombinant CPP-enhanced green fluorescent protein (EGFP). Viability of R1 cells was different between experimental groups depending on the kind of CPP and the concentration of CPP-EGFP. Translocation of CPP-EGFPs into the R1 cells was not detected until 30 min after CPP-EGFPs treatment in all groups. After 1 hr, translocation of pEP-1-EGFP-N was detected, but it could not be detected in the other group. Transfection of pEP-1EGFP-N was independent on its concentration. The time course did not show saturation even after 24 hr in pEP-1-EGFP-N. These results showed that the permeability depended on the kind of CPP and the location of His-tag in the case of examined CPPs, and did not need biological energy. On summary, the efficiency of transfection of CPP-EGFP depends on the CPP sequences but the culture time is not a key factor in transfection for the mouse embryonic stem cell. For the future studies to improve the efficiency of translocation of protein into embryonic stem cells, it is needed to develop modified CPP or mediator. The studies would be very useful to induce the differentiation of embryonic stem cells.

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Study on the Elimination of Heavy Metal Pollutants in the Soil by Vascular Plant in the River Area (하천유역의 관속식물을 이용한 토양내의 중금속 오염원 제거에 관한 연구)

  • 강경홍;김인성
    • Journal of Korea Soil Environment Society
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    • v.1 no.2
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    • pp.23-34
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    • 1996
  • The biological magnification of Pb, Cu, Zn and Fe in vascular plant, Persicaria thunbergii, Trapa japonica and Nymphoides peltata natually grown at the river and riverside and relationship between the concentration of heavy metals in these vascular plants and in the soil were investigated in the Chonju river, Chonju city. The biological magnification values of these metals in P thunbergii were founded as follows ; Pb, Cu, Zn and Fe were 80.4~254.6$mu\textrm{g}$/g, 284.6~688.4$mu\textrm{g}$/g, 635.5~1979.4$mu\textrm{g}$/g and 1160.0~3590.9$mu\textrm{g}$/g respectively. In the case of T. japonica, Pb, Cu, Zn and Fe were 107.8~306.0$mu\textrm{g}$/g, 282.7~963.0p$mu\textrm{g}$/g, 1328.3~3546.$mu\textrm{g}$/g and 656.8~9944.0$mu\textrm{g}$/g and in N. peltata, 140.1~ 193.9$mu\textrm{g}$/g, 187.7~327.3$mu\textrm{g}$/g, 1126.6~1723.6$mu\textrm{g}$/g and 611.7~1914.6$mu\textrm{g}$/g respectively. The amount of the biological magnification of heavy metals in the leaf, stem and root of these plants was generally increased in the sequence of leaf

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A study of growth factors, chondrogenic differentiation of mesenchymal stem cells and cell response by needle size differences in vitro (인간간엽줄기세포의 연골세포 분화 유도 성장인자 및 주사침 크기 차이에 따른 세포반응에 대한 in vitro 연구)

  • Jeongyun Park;Yu Jeong Hwang;Joseph Junesirk Choi;Jin Young Chon;Suk Won Lee
    • Journal of Dental Rehabilitation and Applied Science
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    • v.40 no.1
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    • pp.13-23
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    • 2024
  • Purpose: This aim of this study was to demonstrate growth factors that differentiate human mesenchymal stem cells into chondrocytes and to evaluate cell proliferation enhancement by needle size differences. Materials and Methods: Human mesenchymal stem cells were cultured in chondrogenic medium supplemented with BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1, TGF-β2, TGF-β3 and without growth factors for 14, 21, and 28 days. Then, the expression levels of SOX-5, SOX-6, SOX-9 and FOXO1A were comparatively analyzed. Human mesenchymal stem cells were inoculated into culture dishes using 18, 21, and 26 gauge (G) needles, and cell proliferation was measured after 24, 48, and 72 hours, respectively. Results: In addition to the previously known FGF, IGF-1, and TGFβ1,the BMP family growth factors such as BMP-2, BMP-4, BMP-6, and BMP-7 increased the expression of chondrocyte differentiation genes SOX-5, SOX-6, SOX-9, and FOXO1A. At 48 hours, the 26G group, the smallest needle, showed significant cell proliferation improvement compared to the control group and the 18G group. At 72 hours, the 26G group, the smallest needle, showed significant increase in cell proliferation compared to the control group. Conclusion: Through this study, growth factors with the ability to induce chondrocyte differentiation of human mesenchymal stem cells were investigated, and cell proliferation changes by needle size differences were determined.

Establishment and Characterization of Bone Marrow Mesenchymal Stromal/Stem Cells (MSCs) Derived from ${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) Pig (${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명)

  • Ock, Sun-A;Oh, Keon Bong;Hwang, Seongsoo;Im, Seoki;Kim, Youngim;Park, Jin-Ki
    • 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.

Elimination of Apple stem grooving virus from 'Mansoo' pear (Pyrus pyrifolia L.) by an antiviral agent combined with shoot tip culture (항바이러스제 처리와 경정배양에 의한 배(Pyrus pyrifolia L.) '만수'의 Apple stem grooving virus 무병화)

  • Cho, Kang Hee;Shin, Juhee;Kim, Dae-Hyun;Park, Seo Jun;Kim, Se Hee;Chun, Jae An;Kim, Mi Young;Han, Jeom Hwa;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.391-396
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    • 2016
  • In this study, in vitro-cultured 'Mansoo' pear (Pyrus pyrifolia L.) plants infected with Apple stem grooving virus (ASGV) were used for testing the efficiency of the virus elimination methods. The shoot tips cut from infected plants were treated by thermotherapy ($37^{\circ}C$), cold therapy ($4^{\circ}C$), chemotherapy with ribavirin, and combination of these methods. Treatment periods were 2, 4, and 8 weeks, and concentrations of ribavirin were 20 and $40mg{\cdot}L^{-1}$. The efficiency of ASGV elimination was evaluated by reverse transcription polymerase chain reaction. The shoot survival rate was the highest at 100% after cold therapy, chemotherapy, and combination of two methods, while the rate was the lowest at 33.3% after thermotherapy for 2 weeks. The shoot survival rate after chemotherapy decreased gradually as the treatment period was prolonged. The ASGV elimination rate was the highest at 100% after ribavirin treatment at a concentration of $40mg{\cdot}L^{-1}$ and combination of ribavirin treatment and thermotherapy for 2 weeks, whereas the ASGV elimination rate after cold therapy was the lowest at 16.7%. However, the efficiency of ASGV elimination was enhanced up to 43.3% by the combination of cold therapy and ribavirin treatment. The efficiency of ASGV elimination for all treatments was increased as the treatment period was prolonged. Based on these results, we suggest that ribavirin treatment at a concentration of $20mg{\cdot}L^{-1}$ for 4 weeks or at a concentration of $40mg{\cdot}L^{-1}$ for 2 weeks combined with shoot tip culture was efficient for the elimination of ASGV from pear.

Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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