• 제목/요약/키워드: Cell fusion technique

검색결과 38건 처리시간 0.027초

영상 타원법을 이용한 인간 줄기세포의 굴절률과 두께 분포 연구 (Study on Refractive Index and Thickness of Human Stem Cells by Using Imaging Ellipsometry)

  • 최중규;심우영;이광;김상열;박상욱;제갈원;조현모;조용재
    • 한국광학회지
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    • 제20권1호
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    • pp.53-56
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    • 2009
  • 타원법을 사용하여 인간 중간엽 줄기세포의 광학상수와 두께의 영상정보를 정량적으로 구하였다. 영상 타원계를 이용하여 in vitro 환경에서 세포의 핵(nucleus)과 세포체(cell body)의 타원상수 $\Delta$, $\Psi$를 2차원 영상정보 형태로 구한 다음 각 화소별로 타원상수들을 수치해석적 역방계산하여 굴절률과 두께의 영상정보를 얻었다. 세포의 핵 영역과 세포체 영역에서 줄기세포의 두께와 굴절률 값을 2차원 영상정보 형태로 제시하였다.

Knockdown of Archvillin by siRNA Inhibits Myofibril Assembly in Cultured Skeletal Myoblast

  • Lee, Yeong-Mi;Kim, Hyun-Suk;Choi, Jun-Hyuk;Choi, Jae-Kyoung;Joo, Young-Mi;Ahn, Seung-Ju;Min, Byung-In;Kim, Chong-Rak
    • 대한의생명과학회지
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    • 제13권4호
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    • pp.251-261
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    • 2007
  • A myofiber of skeletal muscle is composed of myofibrils, sarcolemma (plasma membrane), and constameres, which anchor the myofibrils to the sarcolemma. Achvillin is a recently identified F-actin binding muscle protein, co-isolates with dystrophin and caveolin-3 in low-density sarcolemma of striated muscle, and colocalizes with dystrophin at costameres, the specialized adhesion sites in muscle. Archvillin also binds to nebulin and localizes at myofibrillar Z-discs, the lateral boundaries of the sarcomere in muscle. However other roles of archvillin on the dynamics of myofibrillogenesis remain to be defined. The goal of this study is, by using siRNA-mediated gene silencing technique, to investigate the effect of archvillin on the dynamics of myofibrillogenesis in cell culture of a mouse skeletal myogenic cell line (C2C12), where presumptive myoblasts withdraw from the cell cycle, fuse, undergo de novo myofibrillogenesis, and differentiate into mature myotubes. The roles of archvillin in the assembly and maintenance of myofibril and during the progression of myofibrillogenesis induced in skeletal myoblast following gene silencing in the cell culture were investigated. Fluorescence microscopy demonstrated that the distribution of archvillin was changed along the course of myofibril assembly with nebulin, vinculin and F-actin and then located at Z-lines with nebulin. Fluorescence microscopy demonstrated that knockdown of mouse archvillin expression led to an impaired assembly of new myofibrillar clusters and delayed fusion and myofibrillogenesis although the mouse archvillin siRNA did not affect those expressions of archvillin binding proteins, such as nebulin and F-actin. This result is corresponded with that of RT-PCR and western blots. When the perturbed archvillin was rescued by co-transfection with GFP or Red tagged human archvillin construct, the inhibited cell fusion and myotube formation was recovered. By using siRNA technique, archvillin was found to be involved in early stage of myofibrillogenesis. Therefore, the current data suggest the idea that archvillin plays critical roles on cell fusion and dynamic myofibril assembly.

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Microbial Transformation of Aniline to Acetaminophen

  • Lee, Sang-Sup;Jin, Hyung-Jong;Son, Mi-Won
    • Archives of Pharmacal Research
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    • 제15권1호
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    • pp.30-34
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    • 1992
  • In order to obtain acetaminophen, a popular analgesic-antipyretic, through microbial p-hydroxylation and N-acetylation of aniline, various fungi and bacteria were secreened. Among them, Streptomyces species were chosen for strain improvement by the use of interspecific protoplast fusion technique. Two interspecific fused strains were developed between S. rimosus (N-cetylation function) and S. aureofaciens (p-hydroxylation function) and also between S. lividans and S. globisporus. For efficient protoplast fusion and cell wall regeneration, various conditions were examined. In a typical experiment of mixed S rimosus ($pro^- \;his^-$) and S. aureofaciens ($ilv^-$) protoplasts with 40% (w/v) polythylene glycol 3350 (PEG) for 3 min gave $8.3\times10^{-7}$ of fusion frequency. Treatment of mixed S. lividans (pant-) and S. globisporus (leu-) protoplasts with 50% (w/v) PEG for 3 min at $30^\circ{C}$ gave $1.2\times10^{-6}$ of frequency. Among the fused strains, up to 40-50% increase in p-hydroxylation power was observed. To investigate the possibility of plasmid involvement in p-hydroxylation power was observed. To investigate the possibility of plasmid involvement in p-hydroxylation of acetanilide, plasmid curing was attempted. We found that cells treated with acriflavine (at the frequency of 100%) and cells regenerated from protoplsts of S. auroefaciens (2% frequency) lost their p-hydroxylation function.

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돼지 유행성 설사(Porcine Epidemic Diarrhea)의 진단을 위한 면역조직 화학적 기법의 응용 (Application of immunohistochemical technique for diagnosis of porcine epidemic diarrhea)

  • 박남용;조경오
    • 대한수의학회지
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    • 제34권4호
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    • pp.805-813
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    • 1994
  • Immunohistochemical study on the intestinal tissues obtained from the 21 pigs of the 14 terms in Korea in which the clinical and epidemiological features had indicated the possible outbreaks of porcine epidemic diarrhea(PED) was performed using the indirect immunofluorescence test and/or the immunoperoxidase method in order to detect PED viral antigens in the infected cells of the intestines, and histopathological features were described as well. By immunohistochemical analysis, PED viral antigens were detected in the epithelial cells covering the small intestinal villi and recognized slightly in the cells lining the colonic surface epithelium as well. Occasional fluorescence was also seen in a few intestinal crypt epithelium. On light microscopy, the piglets with PED showed marked villous atrophy and fusion, and severe enterocyte degeneration and desquamation. On the other hand, the older pigs more than 4 week old age was mild villous atrophy and fusion, severe villous epithelial cell proliferation, and moderate mononuclear cell infiltration.

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마우스 핵이식란의 동결에 관한 연구 (Studies on cryopreservation of nuclear transplanted mouse embryos)

  • 이병천;조충호;황우석
    • 대한수의학회지
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    • 제33권1호
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    • pp.171-178
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    • 1993
  • The present study was carried out to investigate the developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulse at output voltage of 2.0 kV/cm for $100{\mu}$ sec to induce cell fusion. The recovery rate and developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos was investigated. 1. The recovery rate of nuclear transplanted 2-cell embryos in normal morphology after freezing and thawing was significantly higher in rapid freezing(DMSO 4.5M) than in slow cooling(p<0.01). 2. When the recovered embryos in normal morphology were cultured in vitro, there were no significant differences in the developmental potency to blastocyst between the freezing methods and the concentrations of cryoprotectant. In summary, these experiments have proved that rapid freezing method(DMSO 4.5M) is effective in nuclear transplanted 2-cell mouse embryos. If improved micromanipulation techniques and freezing are combined, nuclear transplantation technique will contribute to the improvement of productivity in livestock animals.

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친환경 기능성 방음벽 (Environmental Functional Soundproof Wall)

  • 김지성;이우미;김일호;김광수
    • 한국도로학회논문집
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    • 제15권5호
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    • pp.65-73
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    • 2013
  • PURPOSES : This study is to suggest future development direction and application of environmental noise barriers as multi-functional soundproof wall. METHODS : Based on the literature review, case study and patent search, research and patent trend were investigated. Patent search was conducted by Patent searching tools, 'Focust'. RESULTS : As environmental noise barriers, Vegetative soundproof wall, photovoltaic soundproof wall, and air-pollution reduction soundproof wall were investigated. First of all, In Korea, Vegetative soundproof wall is being developed mostly as soundproof wall that has vegetation foundation inside, to meet the domestic condition with 23 patent applications. Second, Photovoltaic soundproof wall is being developed mainly with efficiency of photovoltaic system rather than soundproofing. And it is limited to one generation solar cell technology, although Solar cell technology is developing at a rapid pace. On the other hand, for reducing air-pollutant by soundproof wall, a variety of methods are being suggested (filtration, adsorption, and photocatalytic oxidation), and one of them, adsorption are applied for developing air pollution reduction soundproof wall in Korea. CONCLUSIONS: The above soundproof wall is not simple structure, but road facility applied fusion technique. Therefore, as one system, it is difficult to harmonize due to various considerations for design factor. However, if it's possible that a benefits of one system apply to another system, Synergy effect may be created. In the foreseeable future, soundproof wall may be considered as a road system using fusion technique rather than just functional facility. Therefore, substantial studies for applying multi-functional soundproof wall on the road are needed for the future.

Transduction of Tat-Superoxide Dismutase into Insulin-producing MIN6N Cells Reduces Streptozotocin-induced Cytotoxicity

  • Choung, In-Soon;Eum, Won-Sik;Li, Ming-Zhen;Sin, Gye-Suk;Kang, Jung-Hoon;Park, Jin-Seu;Choi, Soo-Young;Kwon, Hyeok-Yil
    • The Korean Journal of Physiology and Pharmacology
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    • 제7권3호
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    • pp.163-168
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    • 2003
  • The reactive oxygen species (ROS) are considered to be an important mediator in pancreatic ${\beta}$ cell destruction, thereby triggering the development of insulin-dependent diabetes mellitus. In the present study, HIV-1 Tat-mediated transduction of Cu,Zn-superoxide dismutase (SOD) was investigated to evaluate its protective potential against streptozotocin (STZ)-induced cytotoxicity in insulin-producing MIN6N cells. Tat-SOD fusion protein was successfully delivered into MIN6N cells in a dose-dependent manner and the transduced fusion protein was enzymatically active for 48 h. The STZ induced-cell destruction, superoxide anion radical production, and DNA fragmentation of MIN6N cells were significantly decreased in the cells pretreated with Tat-SOD for 1 h. Furthermore, the transduction of Tat-SOD increased Bcl-2 and heat shock protein 70 (hsp70) expressions in cells exposed to STZ, which might be partly responsible for the effect of Tat-SOD. These results suggest that an increased of free radical scavenging activity by transduction of Tat-SOD enhanced the tolerance of the cell against oxidative stress in STZ-treated MIN6N cells. Therefore, this Tat-SOD transduction technique may provide a new strategy to protect the pancreatic ${\beta}$ cell destruction in ROS-mediated diabetes.

Expanded Bed Adsorption 크로마토그래피를 사용하여 재조합 E. coli 세포 파쇄액으로부터 내포체 단백질을 직접 재접힘하는 공정 (In Vitro Refolding of Inclusion Body Proteins Directly from E. coli Cell Homogenate in Expanded Bed Adsorption Chromatography)

  • 조태훈;서창우;이은규
    • KSBB Journal
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    • 제16권2호
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    • pp.146-152
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    • 2001
  • rhGH-GST 융합단백질을 사용하여 재조합 대장균 세포 파쇄액으로부터 직접적으로 내포체의 solid-phase 재접힘을 수행할 수 있는 새로운 공정을 개발하였다. 그것은 고체 업자를 제거하는 동시에 초기에 목적딴백질을 흡착 포집할 수 있 는 expanded bed adsorption 크로마토그래피의 장점을 이용한 것이다. 세포 파쇄액 내 용해훤 내포체로부터의 풀린 융합단백질은 expanded bed adsorption 원리에 의해 STREAMLINE DEAE resin에 흡착되고 세포 찌꺼기 등 고체 입자물들은 위 방향 흐름에 의해 효과적으로 제거된다. Urea를 접차적으로 제거함으로써 융합단백질은 고체 matrix 표면에서 재접힘 된 후 염 놓도 구배에 의해 용출된다. 이 새로운 EBA-mediat$\xi$d 재접힘 방법은 응집현상을 획기적으로 줄이고 공정수율윤 향상시킬 뿐 아나라 공정단계 수를 줄일 수 있다. 이 공정은 우리가 알고 있는 한 세계에서 최초로 개발된 공정이며, 현재 single-chain polypeptide, affinity-tagged protein 등과 갈은 다른 행태의 단백질에 EBA를 사용한 재접힘 공정올 적용시키가 위한 연구가 진행되고 있다.

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뉴런온칩 기술: 미세전극칩시스템과 신경세포 패터닝 기술 (Neuron-on-a-Chip technology: Microelectrode Array System and Neuronal Patterning)

  • 남윤기
    • 대한의용생체공학회:의공학회지
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    • 제30권2호
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    • pp.103-112
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    • 2009
  • Neuron-on-a-Chip technology is based on advanced neuronal culture technique, surface micropatterning, microelectrode array technology, and multi-dimensional data analysis techniques. The combination of these techniques allowed us to design and analyze live biological neural networks in vitro using real neurons. In this review article, two underlying technologies are reviewed: Microelectrode array technology and Neuronal patterning technology. There are new opportunities in the fusion of these technologies to apply them in neurobiology, neuroscience, neural prostheses, and cell-based biosensor areas.

Myocardial tissue engineering using electrospun nanofiber composites

  • Kim, Pyung-Hwan;Cho, Je-Yoel
    • BMB Reports
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    • 제49권1호
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    • pp.26-36
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    • 2016
  • Emerging trends for cardiac tissue engineering are focused on increasing the biocompatibility and tissue regeneration ability of artificial heart tissue by incorporating various cell sources and bioactive molecules. Although primary cardiomyocytes can be successfully implanted, clinical applications are restricted due to their low survival rates and poor proliferation. To develop successful cardiovascular tissue regeneration systems, new technologies must be introduced to improve myocardial regeneration. Electrospinning is a simple, versatile technique for fabricating nanofibers. Here, we discuss various biodegradable polymers (natural, synthetic, and combinatorial polymers) that can be used for fiber fabrication. We also describe a series of fiber modification methods that can increase cell survival, proliferation, and migration and provide supporting mechanical properties by mimicking micro-environment structures, such as the extracellular matrix (ECM). In addition, the applications and types of nanofiber-based scaffolds for myocardial regeneration are described. Finally, fusion research methods combined with stem cells and scaffolds to improve biocompatibility are discussed. [BMB Reports 2016; 49(1): 26-36]