• Title/Summary/Keyword: Stem cells

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Hazard Assessment of Green-Wall Plant Campsis grandiflora K. Schum in Urban Areas based on Pollen Morphology and Cytotoxicity (도심 벽면녹화식물 능소화 (Campsis grandiflora K. Schum)의 화분 형태 및 세포독성에 근거한 유해성평가)

  • Kim, Hyun-Jun;So, Soonku;Shin, Chang-Ho;Noh, Hae-ji;Na, Chun-Soo;Lee, You-Mi
    • Korean Journal of Environmental Biology
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    • v.33 no.2
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    • pp.256-261
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    • 2015
  • This study was conducted to assess the influence of hazard pattern in the surface structure of pollen grains of Campsis grandiflora, and cytotoxicity of different part extracts and nectar on RAW264.7 macrophages. The pollen grains were medium sized ($21.8{\mu}m$) with tricolpate aperture type. In equatorial view, the pollens were prolate (P/E=1.8) and the exine pattern was smooth and reticulate. This result contradict with the rumor of having a hook-shaped protuberance that can damage the cornea because we couldn't observed any protuberance on the surface of the outer wall. Furthermore, we investigated the 70% MeOH extracts (flower, leaf, stem) and nectar of C. grandiflora for their cell viability in temporal basis via MTT analysis on RAW264.7 macrophage cells. There was no significant difference in the cytotoxicity among the MeOH extracts and nectar of C. grandiflora after 24 h. However, nectar showed the dosedependent cytotoxicity on RAW264.7 macrophage cells after 48 h.

Comparison of Cell Numbers in Inner Cell Mass vs Trophectoderm and Establishment of ES-like Cells Derived from Day 7 to 9 IVF Bovine Blastocysts (소 체외수정 유래 7~9일령 배반포내에서 내세포괴 대영양배엽세포의 비율 및 배아간세포 확립율의 비교)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.157-165
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    • 1997
  • 포유류 배반포배의 epiblast는 내세포괴에 포함되어 있으며, 이 epiblast cells이 배 및 태아의 생식세포와 일반 체세포로 분화된다 (Beddington, 1986; Lawson 등, 1991). 그런데 조기에 발달된 부화배반포기 배가 지연발생된 부화배반포기 배보다도 많은 epiblast cells을 가지고 있다고 한다(Talbot 등, 1995). 그래서 본 연구에서는 체외수정 유래의 배반포배의 발육속도에 따른 내세포괴/영양배엽세포의 비율 및 배아간세포 확립 효율을 비교하여 발달일령 간에 차이가 있는지를 규명하고자 하였다. 공시한 소의 난포란을 TCM-199에 0.5$\mu\textrm{g}$/ml FSH, 5$\mu\textrm{g}$/ml LH, 10% FBS, 100 units/ml penicilin, 및 100$\mu\textrm{g}$/ml streptomycin을 첨가하여 39$^{\circ}C$, 5% CO2 조건하에서 24시간 동안 체외성숙한 후, 5$\mu\textrm{g}$/ml heparin으로 수정능이 획득된 1$\times$106 sperm/ml의 정자로 체외수정을 유도하였다. 체외수정 후 18~20시간에 과립막세포를 vortexing으로 제거하여 얻은 모든 체외수정란을 3mg/ml BSA, 20${\mu}\ell$/ml NEM amino acids, 40${\mu}\ell$/ml BME amino acids, 10mM glycine 및 1mM alanine이 함유된 CR1aa 배양액에서 BRL 단층세포와 공배양을 실시하였다. 수정후 7, 8 및 9일재 (체외수정일 : 0일)에 확장배반포기까지 발달한 수정란을 이중염색 및 배아간세포의 확립 실험에 공시하였다. 체외수정후 24시간에 분할된 총 1,145개의 수정란이 7, 8 및 9일째에 후기 배반포기까지 각각 222(15.6%), 103(7.2%) 및 52(3.6%)로 발달하여 총 377개 (26.4%)가 발달하였다. 내세포괴/영양배엽세포의 비율은 7일 및 8일째 배반포배에서 각각 47.2$\pm$11.9/95.1$\pm$24.4개 (33/67%) 및 40.3$\pm$12.4/83.3$\pm$26.9개 (33/67%)로서 9일째 배반포배의 19.3$\pm$8.1/62.3$\pm$23.1개 (24/76%) 보다 유의적(P<0.05)으로 높았다. ES-like cells을 확립하기 위하여 후기배반포기 배를 mouse embryonic fibroblast 단층 공배양기에 옮긴 후 5일에 내세포괴의 부착 여부를 판정하고, 10일에 배아간세포의 확립 여부를 판정하였다. 그 결과 7, 8 및 9일째의 배반포기배의 각각 47.7% (82/172), 30.9%(22/71) 및 15.6%(5/32)에서 배아간세포가 확립되었다. 이상의 결과에서 배반포기까지의 발육이 빠른 수정란에서 영양배엽에 대한 내세포괴세포의 비율이 높았고 배아간세포의 확립율도 높다는 사실을 입증할 수 있었으며, 이와 같은 결과에서 체외수정란 유래 배반포배의 질을 결정할 수 있을 것으로 생각된다.

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Histochemical and Cytochemical Studies on the Secondary Xylem Formation during Grafting Process of Pinus thunbergii (접목 과정에서 형성된 신생 2차목부의 조직학적, 세포화학적 연구)

  • Kim, Jong Sik;Lee, Kwang Ho;Kim, Yoon Soo
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.6
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    • pp.1-11
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    • 2006
  • Grafted tissues were investigated using various microscopic techniques. Pinus thunbergii was used as stock and scion and autografted by cleft graft method. Histochemically, grafting processes can be proceeded by four stages: 1) formation of necrotic layer, 2) proliferation of callus, 3) development of neo-cambium from callus, and 4) restoration of new vascular xylem. Necrotic la yer composed of pectin and lignin was gradually degraded during grafting process and disappeared when new union was formed between stock and scion. A large number of starch and lipid bodies in the cytoplasm were also gradually degraded during grafting process and disappeared at the grafting interface. Nucleus and plasmodesmata were not changed. Bubble-like callus was generated from all living parenchyma cells and from the callus. The tracheary elements differentiated from the callus had either reticulate or pit-like thickenings in the secondary walls with bordered pits. Secondary cell wall thickening occurred toward filing to the void parts between reticulated secondary wall. Tracheids formed in the secondary xylem were short with irregular wall thickness. New secondary xylem cells with swirled shapes, which developed in graft union were oriented horizontally and obliquely to axis of the stem.

Transcriptional Regulation of Human Nanog Gene by OCT4 and SOX2 (OCT4와 SOX2에 의한 인간 Nanog 유전자의 전사 조절)

  • Seok, Hyun-Jeong;Kim, Young-Eun;Park, Jeong-A;Lee, Young-Hee
    • Development and Reproduction
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    • v.14 no.2
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    • pp.123-129
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    • 2010
  • Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self-renewal requires many factors such as OCT4, SOX2, and NANOG. It is previously known that OCT4 and SOX2 can bind to NANOG promoter and support Nanog gene expression in mouse ES cells by the detailed studies using the mouse Nanog promoter. Here, we constructed serial deletion mutant promoter-reporter constructs to investigate the human Nanog gene promoter in detail. The highest promoter activity was obtained in the 0.6 kb (-253/+365) promoter-reporter construct which includes the binding sites of OCT4 and SOX2. To further confirm contribution of OCT4 and SOX2 in Nanog gene expression, we introduced site- directed mutation(s) in the OCT4 and/or SOX2 binding sites of the human Nanog promoter 0.6 kb (-253/+365) and checked the influence of the mutation on the promoter activity using human EC cell line NCCIT. Mutation either in OCT4 binding site or SOX2 binding site significantly reduced the activity of Nanog promoter which directly confirmed that OCT4 and SOX2 binding is essential in human Nanog gene expression.

Identification of Cuts-specific Myogenic Marker Genes in Hanwoo by DNA Microarray (DNA Microarray 분석을 통한 한우 부위별 특이 마커 유전자의 발굴)

  • Lee, Eun-Ju;Shin, Yu-Mi;Lee, Hyun-Jeong;Yoon, Du-Hak;Chun, Tae-Hoon;Lee, Yong-Seok;Choi, In-Ho
    • Journal of Animal Science and Technology
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    • v.52 no.4
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    • pp.329-336
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    • 2010
  • Myogenic satellite cells (MSCs) are mononuclear, multipotent progenitors of adult skeletal muscle possessing a capacity of forming adipocyte-like cells (ALC). To identify the skeletal muscle type-specific myogenic and adipogenic genes during MSCs differentiation, total RNA was extracted from bovine MSCs, myotube-formed cell (MFC), and ALC from each of Beef shank, Longissimus dorsi, Deep pectoral, and Semitendinosus. DNA microarray analysis (24,000 oligo chip) comparing MSCs with MFC and ALC, respectively, revealed 135 differentially expressed genes (> 4 fold) among four cuts. Real-time PCR confirmed expression of 29 genes. Furthermore, the whole tissue sample RNAs analysis showed 6 differentially expressed genes in Beef shank. Among which, 1 gene in MSCs, 4 in MFC, and 1 in ALCs were highly expressed. This study will provide an insight for better understanding the molecular mechanism of differentiation of skeletal muscle type-specific MSCs. The identified genes may be used as marker to distinguish skeletal muscle types.

Biological Activity of Recombinant Human Erythropoietin (EPO) In Vivo and In Vitro

  • Park Jong-Ju;Lee Hyen-Gi;Nam In-Suk;Park Hee-Ja;Kim Min-Su;Chung Yun-Hi;Naidansuren Purevjargal;Kang Hye-Young;Lee Poong-Yun;Park Jin-Gi;Seong Hwan-Hoo;Chang Won-Kyong;Kang Myung-Hwa
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.69-73
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    • 2005
  • The hematopoietic growth factor erythropoietin (EPO) is required for the maintenance, proliferation, and differentiation of the stem cells that produce erythrocytes. To analyse the biological activity of the recombinant human EPO (rec-hEPO), we have cloned the EPO cDNA and genomic DNA and produced rec-hEPO in the CHO cell lines. The growth and differentiation of EPO-dependent human leukemic cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of rec-hEPO. MIT assay values were increased by survival of F36E cells at 24h or 72h. The hematocrit and RBC values were increased by subcutaneous injection of 20 IU (in mice) and 100IU(in rats) rec-hEPO. Hematocrit values remarkably increased at $13.2\%$ (in mice) and $12.2\%$ (in rats). The pharmacokinetic behavior with injection of 6 IU of rec-hEPO remained detectable after 24 h in all mice tested. The highest peat appeared at 2h after injection. The long half-life of rec-hEPO is likely to confer clinical advantages by allowing less frequent dosing in patients treated for anemia. These data demonstratethat ree-hEPO produced in this study has a potent activity in vivo and in vitro. The results also suggest that biological activity of ree-hEPO could be remarkably enhanced by genetic engineering that affects the potential activity, including mutants with added oligosaccharide chain and designed to produce EPO-EPO fusion protein.

Inhibition of Adipogenesis in 3T3-Ll Adipocytes with Water and Ethanol Extracts of Cudrania tricuspidata Leaves (꾸지뽕잎(Cudrania tricuspidata) 추출물의 3TS-L1 세포 분화 억제)

  • Do, Gun-Pyo;Lee, Hye-Jin;Do, Jeong-Ryong;Kim, Hyun-Ku
    • Food Science and Preservation
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    • v.18 no.2
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    • pp.244-249
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    • 2011
  • The inhibitory effects of adipogenesis in 3T3-L1 adipocytes with water and ethanol extracts of Curdrania tricuspidata leaves were investigated. The lipid accumulation of ethanol extract from Curdrania tricuspidata leaves at a concentration of 5 mg/mL was reduced by 50% compared with control cells, whereas water extracts reduced by 86%. The survival rate of cell viability test showed 80% in the growth of cells at concentrations of 0.5-5 mg/mL. The contents of DNJ(1-deoxynojirimycin) and rutin were higher in leaves than those of stem. Moreover, DNJ contents were detected in large amounts in ethanol extract ($3921.3{\mu}g/g$) compared with water extract ($2906.3{\mu}g/g$). On the contrary, the rutin contents was detected in large amounts in water extracts compared with ethanol extract. These results indicate that Curdrania tricuspidata leaves extract is potential sources of natural antiobesity agent.

Anti-Diabetic, Alcohol Metabolizing Enzyme, and Hepatoprotective Activity of Acer tegmentosum Maxim. Stem Extracts (산겨릅나무 줄기 추출물의 항당뇨, 알코올 대사 효소 및 간 보호 활성)

  • Cho, Eun Kyung;Jung, Kyung Im;Choi, Young Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1785-1792
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    • 2015
  • This study was carried out to investigate the antidiabetic, alcohol metabolism, anti-inflammatory, and hepatoprotective effects of Acer tegmentosum extracts (ATE). A. tegmentosum has been traditionally used as a folk medicine to treat hepatic disorders. The antioxidative activities of ATE were measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and superoxide (SOD) assay. DPPH radical scavenging and SOD activities of ATE were about 89% and 82.9% at $0.5{\mu}g/mL$, respectively. Alcohol dehydrogenase and acetaldehyde dehydrogenase activities were 118.0% and 177% at 2 mg/mL, respectively. ${\alpha}-Glucosidase$ inhibitory activity of ATE was 75% higher at $50{\mu}g/mL$ and remarkably increased in a dose-dependent manner. Nitric oxide productions in macrophage RAW 264.7 cells stimulated by lipopolysaccharide was reduced to 16.7% by addition of ATE at 1 mg/mL. ATE showed significant protective effects against tacrine-induced cytotoxicity in Hep G2 cells at $100{\mu}g/mL$. Based on our results, we conclude that ATE may be used as a major pharmacological agent and anti-diabetic, anti-hepatitis, and anti-inflammatory remedy.

Highly Expressed Integrin-α8 Induces Epithelial to Mesenchymal Transition-Like Features in Multiple Myeloma with Early Relapse

  • Ryu, Jiyeon;Koh, Youngil;Park, Hyejoo;Kim, Dae Yoon;Kim, Dong Chan;Byun, Ja Min;Lee, Hyun Jung;Yoon, Sung-Soo
    • Molecules and Cells
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    • v.39 no.12
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    • pp.898-908
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    • 2016
  • Despite recent groundbreaking advances in multiple myeloma (MM) treatment, most MM patients ultimately experience relapse, and the relapse biology is not entirely understood. To define altered gene expression in MM relapse, gene expression profiles were examined and compared among 16 MM patients grouped by 12 months progression-free survival (PFS) after autologous stem cell transplantation. To maximize the difference between prognostic groups, patients at each end of the PFS spectrum (the four with the shortest PFS and four with the longest PFS) were chosen for additional analyses. We discovered that integrin-${\alpha}8$ (ITGA8) is highly expressed in MM patients with early relapse. The integrin family is well known to be involved in MM progression; however, the role of integrin-${\alpha}8$ is largely unknown. We functionally overexpressed integrin-${\alpha}8$ in MM cell lines, and surprisingly, stemness features including $HIF1{\alpha}$, VEGF, OCT4, and Nanog, as well as epithelial mesenchymal transition (EMT)-related phenotypes, including N-cadherin, Slug, Snail and CXCR4, were induced. These, consequently, enhanced migration and invasion abilities, which are crucial to MM pathogenesis. Moreover, the gain of integrin-${\alpha}8$ expression mediated drug resistance against melphalan and bortezomib, which are the main therapeutic agents in MM. The cBioPortal genomic database revealed that ITGA8 have significant tendency to co-occur with PDGFRA and PDGFRB and their mRNA expression were up-regulated in ITGA8 overexpressed MM cells. In summary, integrin-${\alpha}8$, which was up-regulated in MM of early relapse, mediates EMT-like phenotype, enhancing migration and invasion; therefore, it could serve as a potential marker of MM relapse and be a new therapeutic target.

Intraannual and Vertical Variations in Rays of Pinus koraiensis and Larix kaempferi Woods Grown in Korea (잣나무와 낙엽송재 방사조직의 연륜 내 및 축방향 변이성)

  • Kim, Nam-Hun
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.1
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    • pp.65-72
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    • 2009
  • Variation of wood rays within a growth ring and along axial direction in Pinus koraiensis and Larix kaempferi, which are important Korean commercial softwoods, was examined to get some useful information for their effective utilization. Ray height in cell number and ray number per $0.25mm^2$ in tangential section were investigated by optical microscopy. The average heights of uniseriate rays in Pinus koraiensis and Larix kaempferi were 6.3 and 9.1 cells, respectively. The average numbers of uniseriate rays per $0.25mm^2$ in Pinus koraiensis and Larix kaempferi were 6.5 and 7.5, respectively. The average heights of fusiform rays were about 20 cells but were not significantly different in both species. From earlywood to latewood within a growth ring, the heights of uniseriate and fusiform rays decreased but their numbers remained almost constant. Along axial direction, the heights of uniseriate rays in both species increased with the increase of stem height, and the numbers of uniseriate rays per $0.25mm^2$ in Pinus koraiensis were nearly constant from 0.2 m to 5.2 m but increased up to 19.2 m beyond 5.2 m. However, a constant pattern in the numbers of uniseriate rays was not observed in Larix kaempferi. The heights of fusiform rays in both species were the highest in middle part of stems but were the lowest in base and top of trees. In conclusion, the ray variation pattern was comparatively clear within a growth ring but not clear along axial direction.