• 제목/요약/키워드: Cell labeling

검색결과 315건 처리시간 0.025초

Characterizing Organelles in Live Stem Cells Using Label-Free Optical Diffraction Tomography

  • Kim, Youngkyu;Kim, Tae-Keun;Shin, Yeonhee;Tak, Eunyoung;Song, Gi-Won;Oh, Yeon-Mok;Kim, Jun Ki;Pack, Chan-Gi
    • Molecules and Cells
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    • 제44권11호
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    • pp.851-860
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    • 2021
  • Label-free optical diffraction tomography (ODT), an imaging technology that does not require fluorescent labeling or other pre-processing, can overcome the limitations of conventional cell imaging technologies, such as fluorescence and electron microscopy. In this study, we used ODT to characterize the cellular organelles of three different stem cells-namely, human liver derived stem cell, human umbilical cord matrix derived mesenchymal stem cell, and human induced pluripotent stem cell-based on their refractive index and volume of organelles. The physical property of each stem cell was compared with that of fibroblast. Based on our findings, the characteristic physical properties of specific stem cells can be quantitatively distinguished based on their refractive index and volume of cellular organelles. Altogether, the method employed herein could aid in the distinction of living stem cells from normal cells without the use of fluorescence or specific biomarkers.

BrdU에 의한 DNA

  • 손우찬;김형진;이영순
    • Toxicological Research
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    • 제7권1호
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    • pp.83-92
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    • 1991
  • Complexities of testis structure and function are emphasized in morphometrical and genotoxic evaluation by statistical analysis. F-344 rats were treated with azinphos methyl, cyclophosphomide, and dichlorvos. And Brdu was injected with intrapertionially before sacrifice. The existence and degree of DNA damage were measured by Brdu labeling index which represented relative amount of Brdu incorporated in DNA, morphometric change was evaluated by the relative length of tubular diameter in circular seminiferous tubules and the number of spermatogonia per Sertoli cell in stage IX seminiferous tubules.

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Mechanistic Insights into the Chemopreventive Action of Phenethy1 Isothiocyanate against Ν-Nitrosobis(2-Oxopropyl) Amine-Induced Carcinogenesis

  • Lee, In-Seon
    • Preventive Nutrition and Food Science
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    • 제1권2호
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    • pp.227-229
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    • 1996
  • The effects of phenthyl isothiocyanate(PEIFTC) on xenobiotic metabolizing enzymes and cell kinetics in the target organs for Ν-nirtosobis(2-oxopropyl) amine(BOP)-tumorigenicity were investigated in female Syrian golden hamsters in order to gain the mechanistic insigths into the chemopreventive action of PEITS against BOP-initiated lung and pancreatic carcinogenesis in hamsters. Hamsters were given BOP subcuteneo-usly(s.c.) and/or PEITC by gavage 2h prior to the BOP treatment. Eight and 24h after the PEITC administration, animals were sacrificed for analyzing P450 isoenzymes, glutathine(GSH), glutathione S-transferase(GST) and cell kinetics. The PEITC pretreatment significantly reduced the hepatic P450 isoenzume levels such as CYP2B1 and DYP1A1 which were significantly increased by the BOP treatment. However, PEITC did not affect the CYP levels in the pancreas and lung. Interestingly, the PEITC pretreatment rather lowered the heparic GST and GSH levels, regradless of BOP administration. Proliferating cell nuclear antigen(PCNA)- labeling indices were dose dependently decreased by PEITC in the pancreas acini and ducts, bronchioles, and renal tubules in which the cell replication was significantly affected by BOP. These results thus suggest that PEITC exerts the chemopreventive effects in hamsters by influencing xenobiotic matabolizing phase I enzymes in the liver and regulating cell kinetics in the target organs.

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In Situ Single Cell Monitoring by Isocyanide-Functionalized Ag and Au Nanoprobe-Based Raman Spectroscopy

  • Lee, So-Yeong;Jang, Soo-Hwa;Cho, Myung-Haing;Kim, Young-Min;Cho, Keun-Chang;Ryu, Pan Dong;Gong, Myoung-Seon;Joo, Sang-Woo
    • Journal of Microbiology and Biotechnology
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    • 제19권9호
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    • pp.904-910
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    • 2009
  • The development of effective cellular imaging requires a specific labeling method for targeting, tracking, and monitoring cellular/molecular events in the living organism. For this purpose, we studied the cellular uptake of isocyanide-functionalized silver and gold nanoparticles by surface-enhanced Raman scattering (SERS). Inside a single mammalian cell, we could monitor the intracellular behavior of such nanoparticles by measuring the SERS spectra. The NC stretching band appeared clearly at ${\sim}2,100cm^{-1}$ in the well-isolated spectral region from many organic constituents between 300 and 1,700 or 2,800 and $3,600cm^{-1}$. The SERS marker band at ${\sim}2,100cm^{-1}$ could be used to judge the location of the isocyanide-functionalized nanoparticles inside the cell without much spectral interference from other cellular constituents. Our results demonstrate that isocyanide-modified silver or gold nanoparticle-based SERS may have high potential for monitoring and imaging the biological processes at the single cell level.

Hesperidin Induces Apoptosis in SNU-668, Human Gastric Cancer Cells

  • Park, Hae-Jeong;Ra, Je-Hyun;Han, Mi-Young;Chung, Joo-Ho
    • Molecular & Cellular Toxicology
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    • 제3권1호
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    • pp.31-35
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    • 2007
  • Hesperidin, known as a flavonoid constituent of citrus, has been known to reduce the proliferation of several cancer cells. We investigated whether hesperidin-induced cell death on SNU-668, human gastric cancer cells. The cytotoxicity of hesperidin on SNU-668 cells was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at the concentration of 1, 10, 50, and 100 ${\mu}M$. Cell viability by hesperidin was 53.18$\pm$2.85% of control value at 100 ${\mu}M$. The cell death by hesperidin showed apoptotic features, which were confirmed using a combination of 4, 6-diamidino-2-phenylindole (DAPI) staining and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. In the apoptosis-regulating genes, treatment of hesperidin decreased mRNA expression of B-cell CLL/lymphoma 2 (BCL2), whereas expression of BCL2-associated X protein (BAX) was increased. The mRNA expression and the activity of caspase3 (CASP3), a major apoptotic factor, was significantly increased by hesperidin treatment. These results suggest that hesperidin could induce apoptosis through CASP3 activation on SNU-668, human gastric cancer cells.

Overexpression of Hypermethylated Homeobox A11 (HOXA11) Inhibits Tumor Cell Growth and Induces Apoptosis in Cervical Cancer

  • Seung-Yul Lee;Tae Jeong Oh;Sungwhan An;Seung-Hoon Lee
    • 한국발생생물학회지:발생과생식
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    • 제28권2호
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    • pp.37-45
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    • 2024
  • This study aimed to elucidate the potential of Homeobox A11 (HOXA11) as a therapeutic target and a diagnostic methylation marker for cervical cancer. Gene expression analysis using cDNA microarray in cervical cancer cell lines revealed significantly reduced expression of the HOXA11 gene. Subsequent investigation of HOXA11 promoter methylation in samples from normal individuals and invasive cervical cancer patients showed over 53.2% higher methylation in cancer scrapes compared to normal scrapes. Furthermore, overexpression of HOXA11, which is downregulated in cervical cancer, strongly suppressed cell growth in cervical cancer cell lines, HeLa and HT3. Additionally, we performed transferase dUTP nick end labeling assay and confirmed that the inhibition of cervical cancer cell proliferation occurred via apoptosis. Mechanistically, overexpression of HOXA11 led to mitochondrial apoptosis characterized by PARP cleavage due to increased c-Myc and enhanced cytochrome C secretion into the cytoplasm. These findings suggest that HOXA11 could potentially serve as a methylation marker for diagnosing cervical cancer and as a novel therapeutic target for its treatment.

In vivo molecular and single cell imaging

  • Hong, Seongje;Rhee, Siyeon;Jung, Kyung Oh
    • BMB Reports
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    • 제55권6호
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    • pp.267-274
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    • 2022
  • Molecular imaging is used to improve the disease diagnosis, prognosis, monitoring of treatment in living subjects. Numerous molecular targets have been developed for various cellular and molecular processes in genetic, metabolic, proteomic, and cellular biologic level. Molecular imaging modalities such as Optical Imaging, Magnetic Resonance Imaging (MRI), Positron Emission Tomography (PET), Single Photon Emission Computed Tomography (SPECT), and Computed Tomography (CT) can be used to visualize anatomic, genetic, biochemical, and physiologic changes in vivo. For in vivo cell imaging, certain cells such as cancer cells, immune cells, stem cells could be labeled by direct and indirect labeling methods to monitor cell migration, cell activity, and cell effects in cell-based therapy. In case of cancer, it could be used to investigate biological processes such as cancer metastasis and to analyze the drug treatment process. In addition, transplanted stem cells and immune cells in cell-based therapy could be visualized and tracked to confirm the fate, activity, and function of cells. In conventional molecular imaging, cells can be monitored in vivo in bulk non-invasively with optical imaging, MRI, PET, and SPECT imaging. However, single cell imaging in vivo has been a great challenge due to an extremely high sensitive detection of single cell. Recently, there has been great attention for in vivo single cell imaging due to the development of single cell study. In vivo single imaging could analyze the survival or death, movement direction, and characteristics of a single cell in live subjects. In this article, we reviewed basic principle of in vivo molecular imaging and introduced recent studies for in vivo single cell imaging based on the concept of in vivo molecular imaging.

Localization of Single Chain Fv Antibodies (scFv) in Transgenic Tobacco Ptants Showing Resistance against Tomato Bushy Stunt Virus

  • Jeun, Y.C.;Boonrod, K.;Nagy, P.;Conrad, U.;Krczal, G.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.75.2-75
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    • 2003
  • To develop an effective protection strategy against tomato bushy stunt virus (TBSV), tobacco plants expressing single-chain Fv antibodies (scFv), were established. A previous had shown that the replication activity of viral replicase was inhibited by the selected scFvs. Moreover, no systemic symptom was found after virus inoculation on leaves of wt N. benthamiana infiltrated with an Agrobacterium suspension resulting i3l expression of the scFvs. However, control plants showed systemic symptoms. In this study the localization of the scFvs within two transgenic plant lines, (CP28H3, CP-P55) was demonstrated using immunogold labelling. The gold particles, indicating the presence of scFv, were mostly found In the cytoplasm of the plant cells including chloroplasts and in the cell walls. However, they were hardly found in the vacuole, nucleoplasm and intercellular spaces. Gold particles often accumulated in either the cytosol or chloroplasts showing a specific labeling, There was no difference in type of gold labeling between both transgenic lines. The localization of the scFv in the cytoplasm further conforms the inhibition of the RNA-dependent RNA polymerase (RdRp) by the selected scFv because it is known that the RdRp is localized to membraneous cytosolic structures.

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AN ELECTRON MICROSCOPIC, RADIOAUTOGRAPHIC STUDY OF ERYTHROPOIESIS IN VITRO

  • MYUNG No Chul
    • 치과방사선
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    • 제17권1호
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    • pp.27-49
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    • 1987
  • Using ³H-proline as a radioactive tracer, the relationship between the ultrastructural differentiation and the site of protein synthesis has been investigated in developing red blood corpuscles. The general ultra-structure of erythropoietic cells in differentiation after 60 minutes of in vitro labeling has confirmed the results from previous investigations by Bessis, M., Thiery, J. and others. In dividing nuclei more than two-thirds of the labeling were present at the interface between heterochromatin and euchromatin. In less differentiated cells most of the grains in interphase cells was localized over the nucleus. As the cells continued to develope beyond a stage where cytoplasmic density was clearly increased over other cell lines in bone marrow, the majority of grains localized over the cytoplasmic area was decreased in more mature cells, as judged by the density of cytoplasm, and the structural changes in mitochondria, Golgi complex and polysomal configurations. These results show; 1) that the cytoplasm of erythroblast series does not change under in vitro conditions employed in the study; 2) that protein synthesis in the nucleus occurs largely at the interface between euchromatin and heterochromatin in active nuclei; and 3) that cytoplasmic synthesis of proteins continues to take place well into the normoblast stage solong as the physically visible polysomes are present in maturing red blood corpuscles.

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Direct radio-iodination of folic acid for targeting folate receptor-positive tumors

  • Huynh, Phuong Tu;Lee, Woonghee;Ha, Yeong Su;Yoo, Jeongsoo
    • 대한방사성의약품학회지
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    • 제4권1호
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    • pp.3-10
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    • 2018
  • The folate receptor (FR) is a promising cell membrane-associated target for nuclear imaging of various cancers (via imaging $FR-{\alpha}$) and potentially also inflammatory diseases (via imaging $FR-{\beta}$), through the use of folic acid-based radioconjugates. However, there have been several drawbacks of previously reported radioconjugates, such as a short half-life of the radiolabel ($^{68}Ga\;t_{1/2}$ 68 min), a complex and time-consuming multistep radiosynthesis, and a high renal uptake of radiolabeled folate derivatives. The goal of this study was to develop an imaging probe by directly labeling folate with radioactive iodine without using an extra prosthetic group. The radiolabeling of folate was optimized using various labeling conditions and the labeled tracers were isolated by high-performance liquid chromatography. The in vitro stability of labeled folate was checked in phosphate-buffered saline and serum. The tumor-targeting efficacy of the probe was also evaluated by biodistribution studies using a murine 4T1 tumor model.