• Title/Summary/Keyword: Expanded stem

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Promising Therapeutic Effects of Embryonic Stem Cells-Origin Mesenchymal Stem Cells in Experimental Pulmonary Fibrosis Models: Immunomodulatory and Anti-Apoptotic Mechanisms

  • Hanna Lee;Ok-Yi Jeong;Hee Jin Park;Sung-Lim Lee;Eun-yeong Bok;Mingyo Kim;Young Sun Suh;Yun-Hong Cheon;Hyun-Ok Kim;Suhee Kim;Sung Hak Chun;Jung Min Park;Young Jin Lee;Sang-Il Lee
    • IMMUNE NETWORK
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    • v.23 no.6
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    • pp.45.1-45.22
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    • 2023
  • Interstitial lung disease (ILD) involves persistent inflammation and fibrosis, leading to respiratory failure and even death. Adult tissue-derived mesenchymal stem cells (MSCs) show potential in ILD therapeutics but obtaining an adequate quantity of cells for drug application is difficult. Daewoong Pharmaceutical's MSCs (DW-MSCs) derived from embryonic stem cells sustain a high proliferative capacity following long-term culture and expansion. The aim of this study was to investigate the therapeutic potential of DW-MSCs in experimental mouse models of ILD. DW-MSCs were expanded up to 12 passages for in vivo application in bleomycin-induced pulmonary fibrosis and collagen-induced connective tissue disease-ILD mouse models. We assessed lung inflammation and fibrosis, lung tissue immune cells, fibrosis-related gene/protein expression, apoptosis and mitochondrial function of alveolar epithelial cells, and mitochondrial transfer ability. Intravenous administration of DWMSCs consistently improved lung fibrosis and reduced inflammatory and fibrotic markers expression in both models across various disease stages. The therapeutic effect of DW-MSCs was comparable to that following daily oral administration of nintedanib or pirfenidone. Mechanistically, DW-MSCs exhibited immunomodulatory effects by reducing the number of B cells during the early phase and increasing the ratio of Tregs to Th17 cells during the late phase of bleomycin-induced pulmonary fibrosis. Furthermore, DW-MSCs exhibited anti-apoptotic effects, increased cell viability, and improved mitochondrial respiration in alveolar epithelial cells by transferring their mitochondria to alveolar epithelial cells. Our findings indicate the strong potential of DW-MSCs in the treatment of ILD owing to their high efficacy and immunomodulatory and anti-apoptotic effects.

Preimplantation Genetic Diagnosis in Inborn Error Metabolic Disorders (유전성 대사질환의 착상전 유전진단)

  • Kang, Inn Soo
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.5 no.1
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    • pp.94-107
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    • 2005
  • Prenatal diagnosis (PND) such as amniocentesis or chorionic villi sampling has been widely used in order to prevent the birth of babies with defects especially in families with single gene disorderor chromosomal abnormalities. Preimplantation genetic diagnosis (PGD) has already become an alternative to traditional PND. Indications for PGD have expanded beyond those practices in PND (chromosomal abnormalities, single gene defects), such as late-onset diseases with genetic predisposition, and HLA typing for stem cell transplantation to affected sibling. After in vitro fertilization, the biopsied blastomere from the embryo is analyzed for single gene defect or chromosomal abnormality. The unaffected embryos are selected for transfer to the uterine cavity. Therefore, PGD has an advantage over PND as it can avoid the risk of pregnancy termination. In this review, PGD will be introduced and application of PGD in inborn error metabolic disorder will be discussed.

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Analysis of Ceramics Using Scanning Electron Microscopy (주사전자현미경을 활용한 세라믹의 분석)

  • Lee, Sujeong
    • Ceramist
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    • v.22 no.4
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    • pp.368-380
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    • 2019
  • A ceramic is used as a key material in various fields. Accordingly, the use of scanning electron microscopy is increased for the purpose of evaluating the reliability and defects of advanced ceramic materials. The scanning electron microscope is developed to overcome the limitations of optical microscopy and uses accelerated electrons for imaging. Various signals such as SE, BSE and characteristic X-rays provide useful information about the surface microstructure of specimens and, the content and distribution of chemical components. The development of electron guns, such as FEG, and the improved lens system combined with the advanced in-lens detectors and STEM-in-SEM system have expanded the applications of SEM. Automated SEM-EDS analysis also greatly increases the amount of data, enabling more statistically reliable results. In addition, X-ray CT, XRF, and WDS, which are installed in scanning electron microscope, have transformed SEM a more versatile analytical equipment. The performance and specifications of the scanning electron microscope to evaluate ceramics were reviewed and the selection criteria for SEM analysis were described.

Establishment of Mouse Embryonic Stem Cell-like Cells from In Vitro Fertilized Embryos (체외수정 생쥐 배아에서의 배아 줄기세포 확립)

  • Shin, Yong-Moon;Park, Yong-Bin;Kim, Hee-Sun;Oh, Sun-Kyung;Chun, Dae-Woo;Suh, Chang-Suk;Choe, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.1-12
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    • 2002
  • Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.

In vitro Neural Cell Differentiation of Genetically Modified Human Embryonic Stem Cells Expressing Tyrosine Hydroxylase (Tyrosine Hydroxylase 유전자가 주입된 인간 배아줄기세포의 체외 신경세포 분화)

  • Shin, Hyun-Ah;Kim, Eun-Young;Lee, Keum-Sil;Cho, Hwang-Yoon;Kim, Yong-Sik;Lee, Won-Don;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.67-74
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    • 2004
  • Objective: This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Materials and Methods: Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA, embryoid bodies (EB, for 4 days) derived from TH#2/MB03 cells were exposed to RA ($10^{-6}M$)/AA ($5{\times}10^{-2}mM$) for 4 days, and were allowed to differentiate in N2 medium for 7, 14 or 21 days. Exp. II) When b-FGF was used, neuronal precursor cells were expanded at the presence of b-FGF (10 ng/ml) for 6 days followed by a final differentiation in N2 medium for 7, 14 or 21 days. Neuron differentiation was examined by indirect immunocytochemistry using neuron markers (NF160 & NF200). Results: After 7 days in N2 medium, approximately 80% and 20% of the RA or b-FGF induced Th#2/MB03 cells were immunoreactive to anti-NF160 and anti-NF200 antibodies, respectively. As differentiation continued, NF200 in RA treated cells significantly increased to 73.0% on 14 days compared to that in b-FGF treated cells (53.0%, p<0.05), while the proportion of cells expressing NF160 was similarly decreased between two groups. However, throughout the differentiation, expression of TH was maintained ($\sim$90%). HPLC analyses indicated the increased levels of L-DOPA in RA treated genetically modified hES cells with longer differentiation time. Conclusion: These results suggested that a genetically modified hES cells (TH#2/MB03) could be efficiently differentiated in vitro into mature neurons by RA induction method.

Analysis of Stromal Cells Developed from Cord Blood CD34+ Cells (제대혈 CD34+ 세포에서 유래된 지지세포의 분석)

  • Ryu, Kyung-Ha;Park, Se-Jin;Kim, Kyung Hyo;Seoh, Ju-Young;Khan, Mohammad;Shin, Hee-Young;Ahn, Hyo-Seop
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.87-94
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    • 2001
  • Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.

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Potential Biological Control of Orobanche by Fungi Isolated from Diseased Specimens in Jordan

  • K. M. Hameed;I. M. Saadoun;Shyab, Zaineb-Al
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.257-263
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    • 2001
  • Species of the genus Orobanche are parasitic flowering plants, holoparasites, which cling to the roots of green plants. Their tiny seeds (200 x $250\mu\textrm{m}$) germinate in response to chemical stimuli produced by host and some non-host plants. Successful contact with their host leads to development of haustoria for obtaining water and food. The shoots above the ground expose flowers and disseminate seeds. Several samples of Orobanche ramosa, O. crenata, O. cernua, and O. egyptiaca were collected from different localities in Jordan. These samples showed one of the following disease symptoms: dry rot at the base of the stem; general deterioration and expanded lesion from base upward; soft tissue maceration of stem; and black rot of flower parts with incomplete maturation of the ovary and seeds. Isolation from diseased stems and seeds was made on three different mycological media. Several fungi were isolated, mainly, Fusarium spp., Alternaria alternata, Rhizoctonia sp., Dendrophora sp., Chaetomium sp., and an ascomycetus fungus with a perithecium. Pathogenicity tests showed that Fusarium spp. and Alternaria alternata attacked healthy living tissue of Orobanche spikes. These fungi caused lesions of black soft rot and complete deterioration within 5-7 days. They also attacked Orobanche seeds, arresting their germination and causing maceration of non-germinated and germinated seeds after 5-7 days of incubation. Meanwhile, Dendrophora sp. and Chaetomium sp. caused limited lesion at first, but were able to colonize the tissue as it aged and senesced. This study showed the presence of a potential endogenous pathogenic fungi in Jordan, which can be investigated as a biological control for Orobanche.

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Identification of Molecular Markers Linked to Lf2 Locus in Soybean

  • Kim Myung-Sik;Park Min-Jung;Jeong Woo-Hyeun;Nam Ki-Chul;Chung Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.2
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    • pp.169-172
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    • 2006
  • Leaflet number of soybean controlled by Lf2 locus is the important trait in photosynthesis and plant type. The objective of this research was to identity molecular markers linked to the lf2 locus. A total of $115F_2$ plants were derived from a cross between normal three-leaflet type Sinpaldalkong (Lf2Lf2) and seven-leaflet mutant type T255 (lf2lf2). All leaflet counts of parents and $F_2$ individual plants were made in the field on fully expanded leaves on the main stem when terminal growth of the main stem had ceased. One-thousand 10-mer oligonucleotide RAPD primers and 664 SSR primers were used. The segregation ratios of 3 : 1 were observed in the $F_2$ population and the Chi-square values strongly suggested that the seven-leaflet was controlled by a single recessive gene. A genetic map was constructed from the 15 segregating markers (9 RAPDs, 5 SSRs, 1 lf2 locus). OPAD03 and OPAI13 RAPD markers were linked to the lf2 locus that controlled seven-leaflet type at a distance of 20.5 and 23.5 cM, respectively. Molecular markers identified in this study linked with lf2 locus will be helpful to locate lf2 locus on the public soybean molecular linkage map and would be useful for tagging the lf2 locus that controls seven-leaflet trait.

Occurrence of Rhizopus Soft Rot on Squash (Cucurbita moschata) Caused by Rhizopus stolonifer in Korea

  • Kwon, Jin-Hyeuk;Kang, Soo-Woong;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • v.16 no.6
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    • pp.321-324
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    • 2000
  • season of 2000. The disease infection usually started from flower, peduncle and young fruits, then moved to flower stalk, stem and leaves. At first, the lesions started with water-soaked, rapidly softened, and then the area gradually expanded. In severely affected film house, the rate of infected fruits reached to 28.6%. Numerous sporangiospores were formed on the diseased fruits, flower stalk, stem and leaves. Most of the sporangiospores were appeare to be rapidly dispersed in the air. The mycelia grew on the surface of host and formed stolons. Colonies on potato dextrose agar were cottony at first brownish black at maturity. Sporangia were 125.3${\times}$294.2 ${\mu}$m. globose or sub-globose with somewhat flattened base. White at first the black, many spored, and are never overhanging. Sporangiophores were 2.7-6.8${\times}$12.9-33.9 ${\mu}$m, smooth-walled, non-septate, light brown, simple, long, arising in groups of 3-5 from stolons opposite rhizoids. Sporangiophores were 8.6-21.1${\times}$6.41-1.7 ${\mu}$m, irregular, round, oval, elongate, angular and brownish-black streaked. Columella were 63.8${\times}$140.4 ${\mu}$m. brownish gray, umberella-shaped when dehisced. The causal organism was identified as Rhizopus stolonifer Lind on the basis of the morphological characteristics of the fungus. Rhizopus soft rot on squash (Cucurbita moschata) caused by the fungi has not been previously reported in Korea.

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Science & Engineering Degrees and Human Resource Element Value Estimation in Technology Jobs : the US Case (기술직에서 이공계학위와 인적자원요소의 가치평가 : 미국사례)

  • Lee, Sae Jae;Lee, Hyun Soo
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.40 no.4
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    • pp.221-229
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    • 2017
  • In the international businesses human resource elements acquired in different countries might have different values in varied industries due to the different quality of education and experiences in the original countries. Using selection models to evaluate expected values in earnings equation of human resource elements such as education and experiences etc. acquired in sending countries, system equations are expanded to examine also the values of science and engineering degrees in technology jobs with selectivity bias correction. This paper used the US census survey data of 2015 on earnings, academic degrees, occupations etc. The US has long maintained the policy of accepting more STEM workers than any other countries and helped maintaining own technological leadership. Assuming per capita GDP gap between the sending country and the US downgrades immigrant human resource quality, it rarely affects occupational selection but depresses earnings on average by two or more years' worth of education. Immigrant quality index in the sense of GDP gap appears to be a valid tool to assess the expected earnings of the worker with. Engineering degrees increase significantly the probability of selecting not only engineering jobs but also general management jobs, as well as increasing the expected earning additionally over nine years'worth of education. Getting a technology job is additionally worth about four years of education. Economics and business degrees are worth additionally almost six years of education but humanities degrees depress expected earnings. Since years after immigration does not very fast enhance earnings capacity, education level and English language ability might be more useful criteria to expect better future earnings by.