• Title/Summary/Keyword: Sf cell

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Preparation and Properties of Silk Fibroin/Alginate Blend Sponges and its Application

  • Kweon, Hae-Yong;Lee, Kwang-Gill;Yeo, Joo-Hong;Woo, Soon-Ok;Han, Sang-Mi;Lee, Yong-Woo;Lee, Jang-Hern;Ham, Tae-Won;Ki, Chang-Seok;Park, Young-Hwan
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.55-56
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    • 2003
  • Silk fibroin (SF) is one of the typical protein polymer produced by silkworm, Bombyx mori. SF has been used as textile fiber and surgical suture fur thousands of years due to its unique gloss, handle, and mechanical properties. Recently, SF has been intensively studied to diverse usage for biotechlological and biomedical fields because of their reproducibility, environmental compatibility, non-toxicity, and biological compatibility. Based on its biocompatibility, the possible uses of regenerated SF have been proposed including substrate for cell culture[1], enzyme immobilization[2], and matrix for drug release[3]. (omitted)

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Production of Recombinant Protein, Human Stem Cell Factor, Using Insect Cell Line

  • Park, Sang-Mi;Kwon, Ki-Sang;Goo, Tae-Won;Yun, Eun-Young;Kang, Seok-Woo;Kim, Sung-Wan;Yu, Kweon;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.37-45
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    • 2009
  • Insect cell cultures have become important tools in the production of biological substances for use in a variety of research, human and veterinary medicine, and pest control applications. These applications often require the introduction of foreign DNA into the cells and have generally used methods originally developed for use with human and other mammalian cell cultures. While these methods can be successfully employed, they are often less efficient with insect cells and frequently involve complex procedures or require specialized equipment. Even when they do work, they may require substantial modification because of differences in the culture medium or growth patterns of insect cells. In this study, We have optimized transfection conditions of Sf9 cell line using insect expression vector pIZT/V5-His which expresses green fluorescent protein effectively. Human stem cell factor (hSCF) is a glycoprotein that plays a key role in hematopoiesis acting both as a positive and negative regulator, often in synergy with other cytokines. It also plays a key role in mast cell development, gametogenesis, and melanogenesis. It can exist in membrane-bound form and in proteolytically released soluble form. As determined by an enzyme-linked immunosorbent assay performed, hSCF level in supernatant averaged 995ng/ml. The human hSCF was partially purified by immunoaffinity chromatography and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The results show that the hSCF has N-linked carbohydrate and corresponds to the soluble form, at or about 223 amino acids in length. The findings suggest functional importance for soluble hSCF in cells.

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Systemic TM4SF5 overexpression in ApcMin/+ mice promotes hepatic portal hypertension associated with fibrosis

  • Joohyeong, Lee;Eunmi, Kim;Min-Kyung, Kang;Jihye, Ryu;Ji Eon, Kim;Eun-Ae, Shin;Yangie, Pinanga;Kyung-hee, Pyo;Haesong, Lee;Eun Hae, Lee;Heejin, Cho;Jayeon, Cheon;Wonsik, Kim;Eek-Hoon, Jho;Semi, Kim;Jung Weon, Lee
    • BMB Reports
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    • v.55 no.12
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    • pp.609-614
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    • 2022
  • Mutation of the gene for adenomatous polyposis coli (APC), as seen in ApcMin/+ mice, leads to intestinal adenomas and carcinomas via stabilization of β-catenin. Transmembrane 4 L six family member 5 (TM4SF5) is involved in the development of non-alcoholic fatty liver disease, fibrosis, and cancer. However, the functional linkage between TM4SF5 and APC or β-catenin has not been investigated for pathological outcomes. After interbreeding ApcMin/+ with TM4SF5-overexpressing transgenic (TgTM4SF5) mice, we explored pathological outcomes in the intestines and livers of the offspring. The intestines of 26-week-old dual-transgenic mice (ApcMin/+:TgTM4SF5) had intramucosal adenocarcinomas beyond the single-crypt adenomas in ApcMin/+ mice. Additional TM4SF5 overexpression increased the stabilization of β-catenin via reduced glycogen synthase kinase 3β (GSK3β) phosphorylation on Ser9. Additionally, the livers of the dualtransgenic mice showed distinct sinusoidal dilatation and features of hepatic portal hypertension associated with fibrosis, more than did the relatively normal livers in ApcMin/+ mice. Interestingly, TM4SF5 overexpression in the liver was positively linked to increased GSK3β phosphorylation (opposite to that seen in the colon), β-catenin level, and extracellular matrix (ECM) protein expression, indicating fibrotic phenotypes. Consistent with these results, 78-week-old TgTM4SF5 mice similarly had sinusoidal dilatation, immune cell infiltration, and fibrosis. Altogether, systemic overexpression of TM4SF5 aggravates pathological abnormalities in both the colon and the liver.

Optimization of fractionation efficiency (FE) and throughput (TP) in a large scale splitter less full-feed depletion SPLITT fractionation (Large scale FFD-SF) (대용량 splitter less full-feed depletion SPLITT 분획법 (Large scale FFD-SF)에서의 분획효율(FE)및 시료처리량(TP)의 최적화)

  • Eum, Chul Hun;Noh, Ahrahm;Choi, Jaeyeong;Yoo, Yeongsuk;Kim, Woon Jung;Lee, Seungho
    • Analytical Science and Technology
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    • v.28 no.6
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    • pp.453-459
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    • 2015
  • Split-flow thin cell fractionation (SPLITT fractionation, SF) is a particle separation technique that allows continuous (and thus a preparative scale) separation into two subpopulations based on the particle size or the density. In SF, there are two basic performance parameters. One is the throughput (TP), which was defined as the amount of sample that can be processed in a unit time period. Another is the fractionation efficiency (FE), which was defined as the number % of particles that have the size predicted by theory. Full-feed depletion mode (FFD-SF) have only one inlet for the sample feed, and the channel is equipped with a flow stream splitter only at the outlet in SF mode. In conventional FFD-mode, it was difficult to extend channel due to splitter in channel. So, we use large scale splitter-less FFD-SF to increase TP from increase channel scale. In this study, a FFD-SF channel was developed for a large-scale fractionation, which has no flow stream splitters (‘splitter less’), and then was tested for optimum TP and FE by varying the sample concentration and the flow rates at the inlet and outlet of the channel. Polyurethane (PU) latex beads having two different size distribution (about 3~7 µm, and about 2~30 µm) were used for the test. The sample concentration was varied from 0.2 to 0.8% (wt/vol). The channel flow rate was varied from 70, 100, 120 and 160 mL/min. The fractionated particles were monitored by optical microscopy (OM). The sample recovery was determined by collecting the particles on a 0.1 µm membrane filter. Accumulation of relatively large micron sized particles in channel could be prevented by feeding carrier liquid. It was found that, in order to achieve effective TP, the concentration of sample should be at higher than 0.4%.

Study of SF6/Ar plasma based textured glass surface morphology for high haze ratio of ITO films in thin film solar cell

  • Kang, Junyoung;Hussain, Shahzada Qamar;Kim, Sunbo;Park, Hyeongsik;Le, Anh Huy Tuan;Yi, Junsin
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.430.2-430.2
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    • 2016
  • The front transparent conductive oxide (TCO) films in thin fill solar cell should exhibit high transparency, conductivity, good surface morphology and excellent light scattering properties. The light trapping phenomenon is limited due to random surface structure of TCO films. The proper control of surface structure and uniform cauliflower TCO films may be appropriate for efficient light trapping. We report light trapping scheme of ICP-RIE glass texturing by SF6/Ar plasma for high roughness and haze ratio of ITO films. It was observed that the variation of etching time, pattern size and Ar flow ratio during ICP-RIE process were important factors to improve the diffused transmittance and haze ratio of textured glass. The ICP-RIE textured glass showed low etching rates due to the presence of metal elements like Al, B, F and Na. The ITO films deposited on textured glass substrates showed the high RMS roughness and haze ratio in the visible wavelength region. The change in surface morphology showed negligible influence on electrical and structural properties of ITO films. The ITO films with high roughness and haze ratio can be used to improve the performance of thin film solar cells.

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Alteration of Stress Fiber in Fibroblastic Reticular Cells via Lymphotoxin β Receptor Stimulation is Associated with Myosin (Lymphotoxin β 수용체를 통한 fibroblastic reticular cell의 stress fiber 변화와 myosin의 연관성)

  • Kim, Min Hwan;Kim, Yeon Hee;Choi, Woobong;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.25 no.5
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    • pp.585-593
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    • 2015
  • Stress fiber (SF) alteration is mediated by cellular receptors, which, upon interaction with the extracellular counterpart, signal to the actin cytoskeleton for remodeling. This association is mediated by a variety of scaffold and signaling factors, which control the mechanical and signaling activities of the interaction site. The heterotrimeric transmembrane lymphotoxin α1β2 (LTα1β2), a member of the tumor necrosis factor (TNF) family of cytokines, including soluble homotrimeric lymphotoxin (LT α), plays an important role in lymphoid tissue architecture. Ligation between LTα1β2 and the lymphotoxin β receptor (LTβR) activates signal-cascade in fibroblastic reticular cells (FRCs). We found LTβR stimulation using an agonistic anti-LTβR antibody alone or combined with LTα or TNFα induced changes in the actin and plasticity of cells. To clarify the involvement of myosin underlying the alteration, we analyzed the effect of myosin light chain kinase (MLCK) with an MLCK inhibitor (ML7), the phosphorylation level of myosin light chains (MLC), and the level of phospho-myosin phosphatase target subunit 1 (MYPT1) after treatment with an agonistic anti-LTβR antibody for cytoskeleton reorganization in FRCs. The inhibition of MLCK activity induced changes in the actin cytoskeleton organization and cell morphology in FRC. In addition, we showed the phosphorylation of MLC and MYPT1 was reduced by LTβR stimulation in cells. A DNA chip revealed the LTβR stimulation of FRC down-regulated transcripts of myosin and actin components. Collectively, these results suggest LTβR stimulation is linked to myosin regarding SF alteration in FRC.

Production of Recombinant Polyhedra Containing Cry1Ac Fusion Protein in Insect Cell Lines

  • Kim, Jae-Su;Choi, Jae-Young;Roh, Jong-Yul;Lee, Han-Young;Jang, Seung-Sik;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.739-744
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    • 2007
  • Insect cell lines and the control of infection for obtaining the maximum amount of polyhedrin-Cry1Ac-polyhedrin fusion protein from Bactrus in monolayer and suspension culture systems were tested. Growth rates of the Trichoplusia ni(High-Five) cell line in both culture systems were better than the other insect cell lines, Spodoptera frugiferda(Sf-9, Sf-21), Trichoplusia ni(Tn5), and Spodoptera exigua(Se301). The expression of the fusion protein in a monolayer culture showed that Se301 cells were 2.3-4.8 times more productive on a per cell basis than the other cell lines. However, in suspension culture, only High-Five cells were productive. High-Five cells infected with Bactrus at a multiplicity of infection(MOI) of 5 and a cell density of $3.0{\times}10^5$ cells per ml were more productive than the other infection condition in a suspension culture suitable for a large-scale production of baculovirus. In conclusion, for the large-scale production of Bactrus in vitro, High-Five cells showing good growth and high productivity are suitable.

Differential Activities of FOXL2 and Its Mutants on SF-1-Induced CYP19 Transcriptional Activation (SF-1을 매개한 CYP19의 전사활성에 미치는 FOXL2 야생형과 돌연변이형의 차별적 영향)

  • Park, Mi-Ra;Kim, Ah-Young;Na, Soon-Young;Kim, Hong-Man;Lee, Kang-Seok;Bae, Jee-Hyeon;Ko, Jeong-Jae
    • Development and Reproduction
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    • v.14 no.2
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    • pp.91-97
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    • 2010
  • FOXL2 is a winged-helix/forkhead (FH) domain transcription factor, and mutations in FOXL2 gene are responsible for blepharophimosis-ptosis-epicanthus inversus syndrome (BPES). BPES is an autosomal dominant genetic disease. BPES type I patients exhibit both premature ovarian failure (POF) and eyelid malformation, while only the eyelid defect is observed in BPES type II. FOXL2-null ovaries showed a blockage of granulosa cell differentiation, suggesting that FOXL2 plays an essential role for proper ovarian folliculogenesis. Previously, we screened for FOXL2-interacting proteins and identified steroidogenic factor-1 (SF-1) which is known to be required for gonad development and transactivates steroidogenic enzymes including CYP19. In the present study, we demonstrated that FOXL2 transactivates CYP19 and stimulated the transcriptional activation of CYP19 induced by SF-1. In contrast, FOXL2 mutants found in BPES type I and II exhibited compromised abilities to enhance CYP19 induction mediated by SF-1. Thus, this study provides a functional difference between wild-type FOXL2 and its mutants which may aid to understand pathophysiology of BPES elicited by FOXL2 mutations.

Effects of Schisandrae Fructus 70% Ethanol Extract on Proliferation and Differentiation of Human Embryonic Neural Stem Cells (오미자 70% 에탄올 추출물의 신경줄기세포 증식과 분화에 미치는 영향)

  • Baral, Samrat;Pariyar, Ramesh;Yoon, Chi-Su;Yun, Jong-Min;Jang, Seok O;Kim, Sung Yeon;Oh, Hyuncheol;Kim, Youn-Chul;Seo, Jungwon
    • Korean Journal of Pharmacognosy
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    • v.46 no.1
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    • pp.52-58
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    • 2015
  • Neural stem cells (NSCs), with self-renewal and neuronal differentiation capacity, are a feasible resource in cell-based therapies for various neurodegenerative diseases and neural tissue injuries. In this study, we investigated the effects of Schisandrae Fructus (SF) on proliferation and differentiation of human embryonic NSCs. Treatment with 70% ethanol extract of SF increased the viability of NSCs derived from human embryonic stem cells, which was accompanied by increased mRNA expression of cyclin D1. Whereas 70% ethanol extract of SF also decreased the mRNA expression of nestin, it increased class III ${\beta}$-tublin (Tuj-1) and MAP2 in both growth and differentiation media. Lastly, we found increased mRNA expression of BDNF in SF-treated NSCs. In conclusion, our study demonstrates for the first time that SF induced proliferation and neuronal differentiation of NSCs and increased mRNA expression of BDNF, suggesting its potential as a regulator of NSC fate in NSC-based therapy for neuronal injuries from various diseases.

Mixture of Corni Fructus and Schisandrae Fructus improves testosterone-induced benign prostatic hyperplasia through regulating 5α-reductase 2 and androgen receptor

  • Hyun Hwangbo;Min Yeong Kim;Seon Yeong Ji ;Beom Su Park;TaeHee Kim;Seonhye Yoon;Hyunjin Kim;Sung Yeon Kim ;Haeun Jung;Taeiung Kim;Hyesook Lee;Gi-Young Kim;Yung Hyun Choi
    • Nutrition Research and Practice
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    • v.17 no.1
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    • pp.32-47
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    • 2023
  • BACKGROUND/OBJECTIVES: Benign prostatic hyperplasia (BPH) characterized by an enlarged prostate gland is common in elderly men. Corni Fructus (CF) and Schisandrae Fructus (SF) are known to have various pharmacological effects, including antioxidant and anti-inflammatory activities. In this study, we evaluated the inhibitory efficacy of CF, SF, and their mixture (MIX) on the development of BPH using an in vivo model of testosterone-induced BPH. MATERIALS/METHODS: Six-week-old male Sprague-Dawley rats were randomly divided into seven groups. To induce BPH, testosterone propionate (TP) was injected to rats except for those in the control group. Finasteride, saw palmetto (SP), CF, SF, and MIX were orally administered along with TP injection. At the end of treatment, histological changes in the prostate and the level of various biomarkers related to BPH were evaluated. RESULTS: Our results showed that BPH induced by TP led to prostate weight and histological changes. Treatment with MIX effectively improved TP-induced BPH by reducing prostate index, lumen area, epithelial thickness, and expression of BPH biomarkers such as 5α-reductase type 2, prostate-specific antigen, androgen receptor, and proliferating cell nuclear antigen compared to treatment with CF or SF alone. Moreover, MIX further reduced levels of elevated serum testosterone, dihydrotestosterone, and prostate-specific antigen in BPH compared to the SP, a positive control. BPH was also improved more by MIX than by CF or SF alone. CONCLUSIONS: Based on the results, MIX is a potential natural therapeutic candidate for BPH by regulating 5α-reductase and AR signaling pathway.