• 제목/요약/키워드: Cell manipulation

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암세포주와 공동 배양된 인간 지방 조직 유래 중간엽 줄기 세포의 유전자 발현 분석 (Analysis of Global Gene Expression Profile of Human Adipose Tissue Derived Mesenchymal Stem Cell Cultured with Cancer Cells)

  • 김종명;유지민;배용찬;정진섭
    • 생명과학회지
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    • 제21권5호
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    • pp.631-646
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    • 2011
  • 중간엽 줄기 세포는 다분화능을 가지고 있으며 골수, 지방, 태반, 치아속질, 윤활막, 편도 및 가슴샘 등 인체의 다양한 조직에서 분리된다. 중간엽 줄기세포는 조직의 항상성을 조절하며 다분화능, 분리와 조작의 용이함, 암세포로의 화학주성 및 면역 반응 조절 등의 특징을 가지고 있어서 재생 의학, 암 치료 및 식대주 질환(GVHD) 등에 이용할 수 있는 세포치료제로 주목 받고 있다. 하지만 주위 세포와 조직을 지지하고 조절하는 특징과 관련하여 중간엽 줄기세포가 혈관 생성을 촉진하고 성장인자를 분비하며 암세포를 공격하는 면역 반응을 억제함으로써 암의 진행을 촉진시킨다는 사실 또한 보고 되고 있다. 이러한 사실들로 인해 중간엽 줄기세포의 임상 적용이 제한되고 있다. 본 연구에서는 어떠한 기전을 통해서 중간엽 줄기세포가 암의 진행을 촉진하는 지지 세포로 기능하는지를 밝히기 위해서 인체 지방 조직에서 유래한 중간엽 줄기세포를 두 개의 암세포주(H460, U87MG)와 각각 공동 배양하고 microarray를 이용해서 암세포와 공동 배양되지 않은 중간엽 줄기세포와 유전자의 발현을 비교하였다. 두 암세포주와 공동배양에서 공통적으로 2배 이상 차이 나는 유전자를 DAVID (Database for Annotation, Visualization and Integrated Discovery)와 PANTHER (Protein ANalysis THrough Evolutionary Relationships)를 이용해 분석하였으며 생물학적 과정, 분자적 기능, 세포의 구성 성분, 단백질의 종류, 질병과 인체 조직 그리고 신호전달에 관련된 정보를 획득하였다. 이를 통해서 암세포는 중간엽 줄기세의 분화, 증식, 에너지 대사, 세포의 구조 및 분비기능을 조절하여 유전자의 발현 양상을 암 연관 섬유모세포(cancer associated fibroblast)와 유사한 세포로 변형 시킨다는 사실을 알 수 있었다. 본 연구의 결과는 중간엽 줄기세포를 이용한 임상 치료제의 효과와 안정성을 개선하는데 응용될 수 있을 것이다.

Enhanced Production of Valuable Bioactive Metabolites in Submerged Cultures of Medicinal Mushroom Ganoderma lucidum by Manipulation of Oxygen Supply

  • Zhong, Jian-Jiang;Fang, Qing-Hua;Tang, Ya-Jie
    • Journal of Plant Biotechnology
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    • 제4권3호
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    • pp.109-115
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    • 2002
  • Submerged cultures of Ganoderma lucidum, a valuable mushroom in traditional Chinese medicine, were used for production of bioactive Banoderic acids and Ganoderma polysaccharides. The significant effects of oxygen supply were demonstrated in both shake flasks and bioreactors. By changing the medium loading volume in a shake flask, a different value of initial volumetric oxygen transfer coefficient ($K_L$a) was obtained, and a higher $K_L$a value led to a higher biomass density and a higher productivity of both intracellular polysaccharide and ganoderic acid. In a stirred bioreactor, at an initial $K_L$a of 78.2 $h^{-1}$, a maximal cell concentration of 15.6 g/L by dry weight was obtained, as well as a maximal intracellular polysarcharide (IPS) production of 2.2 g/L and its maximal productivity of 220 mg/(L.d). An increase of initial $K_L$a led to a higher production and productivity of GA, and the GA production and productivity at an initial $K_L$a of 96.0 $h^{-1}$ was 1.8-fold those at an initial $K_L$a of 16.4 $h^{-1}$. The fundamental information obtained in this study may be useful for efficient large-scale production of these valuable bioactive products by the submerged cultures.

Recent Progress in Biotechnology-based Gene Manipulating Systems to Produce Knock-In/Out Mouse Models

  • Lee, Woon Kyu;Park, Joong Jean;Cha, Seok Ho;Yun, Cheol-Heui
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권5호
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    • pp.745-753
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    • 2008
  • Gene-manipulated mice were discovered for the first time about a quarter century ago. Since then, numerous sophisticated technologies have been developed and applied to answer key questions about the fundamental roles of the genes of interest. Functional genomics can be characterized into gain-of-function and loss-of-function, which are called transgenic and knock-out studies, respectively. To make transgenic mice, the most widely used technique is the microinjection of transgene-containing vectors into the embryonic pronucleus. However, there are critical drawbacks: namely position effects, integration of unknown copies of a foreign gene, and instability of the foreign DNA within the host genome. To overcome these problems, the ROSA26 locus was used for the knock-in site of a transgene. Usage of this locus is discussed for the gain of function study as well as for several brilliant approaches such as conditional/inducible transgenic system, reproducible/inducible knockdown system, specific cell ablation by Cre-mediated expression of DTA, Cre-ERTM mice as a useful tool for temporal gene regulation, MORE mice as a germ line delete and site specific recombinase system. Techniques to make null mutant mice include complicated steps: vector design and construction, colony selection of embryonic stem (ES) cells, production of chimera mice, confirmation of germ line transmission, and so forth. It is tedious and labor intensive work and difficult to approach. Thus, it is not readily accessible by most researchers. In order to overcome such limitations, technical breakthroughs such as reporter knock-in and gene knock-out system, production of homozygous mutant ES cells from a single targeting vector, and production of mutant mice from tetraploid embryos are developed. With these upcoming progresses, it is important to consider how we could develop these systems further and expand to other animal models such as pigs and monkeys that have more physiological similarities to humans.

Polyunsaturated Fatty Acids, Lipid Peroxidation and Antioxidant Protection in Avian Semen - Review -

  • Surai, P.F.;Fujihara, N.;Speake, B.K.;BrilIard, J-P.;Wishart, G.J.;Sparks, N.H.C.
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권7호
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    • pp.1024-1050
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    • 2001
  • Avian spermatozoa are characterised by high concentrations of polyunsaturated fatty acids (PUFAs), in particular docosatetraenoic (DTA, 22:4n-6) and arachidonic (AA, 20:4n-6) acids. As a result they are vulnerable to lipid peroxidation, which is considered to be an important factor of male infertility. Antioxidant systems are expressed in spermatozoa and seminal plasma and build three major levels of antioxidant defense. The first level is based on the activity of superoxide dismutase (SOD) which is, in conjunction with glutathione peroxidase (GSH-Px), catalase and metal-binding proteins, responsible for prevention of free radical formation. The second level of defence is responsible for prevention and restriction of chain reaction propagation and includes chain-breaking antioxidants such as vitamin E, ascorbic acid, glutathione and some others. The third level of antioxidant defence deals with damaged molecules, repairing or removing them from the cell and includes specific enzymes such as lipases, proteases, DNA repair enzymes etc. In the review, profiles of PUFAs and the two first lines of antioxidant defence in avian spermatozoa are characterised. Dietary manipulation of the breeder's diet (PUFA, vitamin E and selenium) as an effective means of modulating fatty acid composition and antioxidant system is also considered. Antioxidant properties of seminal plasma and efficiencies of inclusion of antioxidants into semen diluents are also characterised.

`Ailsa Craig` 토마토 캘러스의 형태학적 특성과 식물체 재분화에 미치는 영향 (Morphological Characterization of 'Ailsa Craig' Tomato Callus and Effect on Plant Regeneration)

  • 성은수;왕명현
    • Applied Biological Chemistry
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    • 제50권4호
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    • pp.253-256
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    • 2007
  • `Ailsa Craig` 토마토 품종의 기내배양시 신초재분화의 최적조건을 밝히고자 실험하였다. 본 실험에서 캘러스 형성 및 신초 재분화에 사용된 배지조성은 6가지 조건으로 하였다. 기본 MS 배지에 zeatin 2 mg/l, zeatin 2 mg/l+IAA 0.1 mg/l, zeatin 2 mg/l+IAA 0.5 mg/l, zeatin 4 mg/l, 4 mg/l+IAA 0.1 mg/l, zeatin 4 mg/l+1AA 0.5 mg/l을 첨가하여 캘러스 및 신초 재분화율의 조건별 차이를 살펴 보았다. 그 결과 zeatin 2 mg/l를 단독 배양한 조건에서 신초 재분화율이 가장 양호하였으며, 캘러스 형성률에 있어서는 큰 타이를 보여주지 못했다. 또한 조건별로 기내상태에 있는 캘러스의 형태적인 차이를 살펴보기 위하여 SEM을 이용하여 관찰하였다. 캘러스 조직의 표면을 관찰한 결과, zeatin 2 mg/l를 단독 배양한 조건에서 완전한 신초로 재분화 하기 위한 구형의 모형이 관찰되었으며, 나머지 조건의 캘러스는 쭈글쭈글한 형태의 캘러스가 확인되었다. 이 같은 결과들로 볼 때, 토마토 'Ailsa Craig'의 조직배양시 캘러스 모양이 찌그러지지 않는 완전한 구형의 캘러스 상태에서 신초재분화율이 더 높아지는 것으로 나타났다.

하이브리드 슈퍼커패시터의 음극 및 양극 설계에 따른 전기화학적 거동 (Electrochemical Behavior Depending on Designed-Anode and Cathodes of Hybrid Supercapacitors)

  • 신승일;이병관;하민우;안건형
    • 한국재료학회지
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    • 제29권12호
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    • pp.774-780
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    • 2019
  • The performance of Li-ion hybrid supercapacitors (asymmetric-type) depends on many factors such as the capacity ratio, material properties, cell designs and operating conditions. Among these, in consideration of balanced electrochemical reactions, the capacity ratio of the negative (anode) to positive (cathode) electrode is one of the most important factors to design the Li-ion hybrid supercapacitors for high energy storing performance. We assemble Li-ion hybrid supercapacitors using activated carbon (AC) as anode material, lithium manganese oxide as cathode material, and organic electrolyte (1 mol L-1 LiPF6 in acetonitrile). At this point, the thickness of the anode electrode is controlled at 160, 200, and 240 ㎛. Also, thickness of cathode electrode is fixed at 60 ㎛. Then, the effect of negative and positive electrode ratio on the electrochemical performance of AC/LiMn2O4 Li-ion hybrid supercapacitors is investigated, especially in the terms of capacity and cyclability at high current density. In this study, we demonstrate the relationship of capacity ratio between anode and cathode electrode, and the excellent electrochemical performance of AC/LiMn2O4 Li-ion hybrid supercapacitors. The remarkable capability of these materials proves that manipulation of the capacity ratio is a promising technology for high-performance Li-ion hybrid supercapacitors.

Assessment of the effects of virus-mediated limited Oct4 overexpression on the structure of the hippocampus and behavior in mice

  • Sim, Su-Eon;Park, Soo-Won;Choi, Sun-Lim;Yu, Nam-Kyung;Ko, Hyoung-Gon;Jang, Deok-Jin;Lee, Kyung-Min;Kaang, Bong-Kiun
    • BMB Reports
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    • 제44권12호
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    • pp.793-798
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    • 2011
  • Recently, pluripotency induction or cellular reprogramming by introducing critical transcription factors has been extensively studied, but has been demonstrated only in vitro. Based on reports that Oct4 is critically involved in transforming neural stem cells into pluripotent cells, we used the lentiviral vector to introduce the Oct4 gene into the hippocampal dentate gyrus (DG) of adult mice. We examined whether this manipulation led to cellular or behavioral changes, possibly through processes involving the transformation of NS cells into pluripotent cells. The Oct4 lentivirus-infused group and the green fluorescent protein lentivirus-infused group showed a similar thickness of the DG and a comparable level of synaptophysin expression in the DG. Furthermore, our behavioral analyses did not show any differences between the groups concerning exploratory activity, anxiety, or memory abilities. This first trial for pluripotency induction in vivo, despite negative results, provides implications and information for future studies on in vivo cellular reprogramming.

Studies on the Viability of Frozen Removed Seminal Plasma by Saline(RSP-S) and Tris-buffer(RSP-T) Semen of Small Spcies Dogs

  • Kim, S. K.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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    • pp.57-57
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    • 2001
  • This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, RSP-S and RSP-T semen and fractional semen of small size dogs, and the effect of temperature and preservation time and cryoproservation on motility of whole and RSP-S and RSP- T semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. The volume per ejaculate semen, sperm of concentration and motility and abnormal sperm rate of 1st fractional semen were 0.65±0.09㎖, 4.52±0.35×10/sup 6/ cells/㎖, 15.64±3.85% and 5.50±0.62%. Also, 2nd fractional semen were 1.25±0.20㎖, 3.35±0.48×10/sup 6/cells/㎖, 96.25±4.65% and 4.24±0.46%. And 3rd fractional semen were 1.45±0.21㎖, 3.85±0.52×10/sup 6/cell/㎖, 92.82±4.24% and 4.66±0.58%, respectively. 2. The sperm of concentration and motility and abnormal sperm rates of whole, RSP-S and RSP-T semen were 5.45±0.82×10/sup 6/ cells/㎖, 95.55±4.65%, 4.58±0.45% and 4.82±0.36×10/sup 6/cells/㎖, 90.10±3.42%, 6.48±0.68% and 4.55±0.45× 10/sup 6/cells/㎖, 93.25±3.85%, 4.82±0.58%, respectively. 3. The motility of whole, RSP-S and RSP-T semen were higher at 4℃ than at 38℃. When preservation temperature was at 4℃, survival rates of RSP-S and RSP-T sperm were 97.54%-6.25% at 1-72 hrs, 97.40%-5.62% at 1-100 hrs, respectively. 4. The survival rates of slow and rapid frozen 2nd fraction, RSP-S and RSP-T semen were 67.3±4.45%, 88.8±4.46% and 46.4±3.84%, 74.4±4.20%, respectively. Survival rates was significantly higher in frozen RSP-S and RSP-T semen than that in control group(8.5±2.12%).

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Conjugated Linoleic Acid (CLA) Production in the Rumen -Roles of Butyrivibrio fibrisolvens A38

  • Kim, Dae-Ok;Kim, Tae-Wan;Heo, Ho-Jin;Imm, Jee-Young;Hwang, Han-Joon;Oh, Sejong;Kim, Young-Jun
    • 한국축산식품학회지
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    • 제24권3호
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    • pp.303-309
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    • 2004
  • Conjugated linoleic acid (CLA) is currently under intensive investigation due to its health benefits. A great deal of interest has been paid to the possible health-promoting roles of CLA, but there are not many studies available on the mechanism of CLA production by ruminal microorganisms. CLA is produced as an intermediate of the characteristic biohydrogenation process of linoleic acid(LA) in the rumen and its production has direct relationship to numerous environmental factors including particle association, substrate concentration, forage-to-grain ratio, pH, ionopore, bacterial cell density, etc. Some of these factors were known to affect hydrogenating activities of Butyrivibrio fibrisolvens A38 which is an active rumen bacterium in CLA production. Dairy cow is a main source of CLA, and its level could be increased by dietary manipulation changing the physiological environment of rumen bacteria such as B. fibrisolvens A38. Therefore, the effects of various factors on. ruminal biohydrogenation should be carefully considered to optimize not only CLA production, but also other fatty acid metabolism, both of which are directly affecting nutritional quality and functionality of dairy products. In this review, the relationship between various environmental factors and ruminal CLA production is discussed focusing on the CLA production of B. fibrisolvens A38.

Nutritional Value of Candida utitis for Rotifer and Larval Flounder Paralichthys olivaceus

  • Kim Hae Young;Kim Joong Kyun;Park Kyong-Joo;Bae Jean Hee;Hur Sung Bum
    • Fisheries and Aquatic Sciences
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    • 제8권4호
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    • pp.235-242
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    • 2005
  • Baker's yeast, Saccharomyces cerevisiae, has been widely used as a food organism for rotifers used in the larval production of marine fish. However, the nutritional value of the yeast is relatively poor compared with that of the marine alga Chlorella. We examined the nutritional value of another yeast, Candida utilis, and whether its food value could be increased through manipulation such as a cell wall treatment. Candida utilis and S. cerevisiae and their manipulated varieties were assessed with regard to the growth and nutrition of the rotifer Brachianus plicatilis. Larvae of the flounder Paralichthys alivaceus were cultured with rotifers fed on the yeast species, and the dietary value of the rotifers for the larvae was examined. Rotifers that were fed C. utilis grew faster than those provided with S. cerevisiae. Rotifers grew slightly faster on manipulated yeast than on non-manipulated yeast varieties. Of the two yeast species, C. utilis had better dietary value for rotifers. Flounder larvae cultured with rotifers that had fed on C. utilis displayed better growth and survival ($\%$) than did those cultured with rotifers that had fed on S. cerevisiae. Although the manipulated variety of C. utilis was better than the non-manipulated variety in terms of rotifer growth, the flounder larvae survived ($\%$) and grew better when they were fed rotifers that had eaten non-manipulated C. utilis. However, the nutritional value of this yeast species was still lower than that of Chlorella.