• Title/Summary/Keyword: live cell

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Studies on the artificial infection and the hematological change with Fasciola hepatica metacercariae in rats (흰쥐에 대(對)한 간질피낭유충(肝蛭被囊幼蟲)의 인공감염(人工感染)과 혈액상(血液像)에 관(關)한 연구(硏究))

  • Kang, Seung-won
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.549-557
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    • 1994
  • This study was done to find the method of the extermination of Fasciola hepatica matacercariae. And the artificial infection was carried out with 30 metacercarae exposed to 5% ammonia water and not-exposed to 5% ammonia water. Serial determinations of live weight, red blood cell, hemoglobin, packed cell volume, and eosinophils were performed in rats at 7 days interval for 16 weeks after infection (WAI). Recovery of worm burden and microscopic findings of livr was performed in rats at 10 WAI. The results in this work were summarized as follows; 1. Fasciola spp metacercariae exposed to 5% ammonia water have lost their ability of infection. 2. In teh exposed group, the mean of worm recovered was 2.25 and the common bile duct was swelling up to 0.71cm in diameter. 3. The value of live weight was different in two groups as the not-exposed group and the exposed group were 321.28, 384.38 at 10 WAI, respectively. 4. In the not-exposed group, at 7 WAI, hemoglobin at 5 WAI and packed cell volume at 7 WAI wre minimally decreased to $5.84{\times}10^{-6}/mm^3$, 11.53g/dl and 43.2%, respectively. But those three values were slowly increased at 10 WAI. Rercent cosinophil was increased to 12.2% at 4 WAI and slightly decreased to 7.9% at 10 WAI. But there are no stastistical singnificance between the exposed group and the normal control group. 5. In histolgical findings in the not-exposed group, the dilated common bile ducts and intrahepatic bile ducts showed distinct hyperplasia of the epithelium. Lymphocytes and eosinophils were infilterated around the bile ducts. The hepatic cells and Kupffer cells showed swollen appearance.

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Real-time FRET imaging of cytosolic FAK signal on microwavy patterned-extracellular matrix (ECM) (미세파상 패턴 ECM 에서 세포질 FAK 신호의 실시간 FRET 이미징)

  • Suh, Jung-Soo;Jang, Yoon-Kwan;Kim, Tae-Jin
    • Journal of Biomedical Engineering Research
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    • v.40 no.1
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    • pp.1-6
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    • 2019
  • Human mesenchymal stem cells (hMSC) are multipotent stromal cells that have great potential to differentiate into a variety of cell types such as osteocytes, chondrocytes, and myocytes. Although there have been many studies on their clinical availability, little is known about how intracellular signals can be modulated by topographic features of the extracellular matrix (ECM). In this study, we investigated whether and how microwavy-patterned extracellular matrix (ECM) could affect the signaling activity of focal adhesion kinase (FAK), a key cellular adhesion protein. The fluorescence resonance energy transfer (FRET)-based FAK biosensor-transfected cells are incubated on microwavy-patterned surfaces and then platelet derived growth factor (PDGF) are treated to trigger FAK signals, followed by monitoring through live-cell FRET imaging in real time. As a result, we report that PDGF-induced FAK was highly activated in cells cultured on microwavy-patterned surface with L or M type, while inhibited by H type-patterned surface. In further studies, PDGF-induced FAK signals are regulated by functional support of actin filaments, microtubules, myosin-related proteins, suggesting that PDGF-induced FAK signals in hMSC upon microwavy surfaces are dependent on cytoskeleton (CSK)-actomyosin networks. Thus, our findings not only provide new insight on molecular mechanisms on how FAK signals can be regulated by distinct topographical cues of the ECM, but also may offer advantages in potential applications for regenerative medicine and tissue engineering.

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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    • v.44 no.11
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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.

Ultrarapid Freezing of Biopsied Mouse Embryos at the 4-cell Stage (할구 한 개가 제거된 생쥐 4세포기 수정란의 초급속동결)

  • 강만종;이철상;한용만;유대열;이경광
    • Journal of Embryo Transfer
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    • v.7 no.2
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    • pp.81-88
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    • 1992
  • Cryopreservation of mouse embryos biopsied at 4-cell stage was investigated by ultrarapid freezing. Four-cell embryos were obtained from ICR mice on 55h after hCG injection. Zona pellucida of the embryos were partially dissected with a cutting pipet, and then single blastomeres were biopsied from the embryos followed by incubation in $Ca^2$+ and $Mg^2$+-free M16 medium for 30min. Biopsied embryos cultured for lh or 15h were frozen by ultrarapid freezing method using 3M DMSO or 5M glycerol as a cryoprotectant, respectively. The developmental rate of biopsied embryos after ultrarapid freezing and thawing to blastocysts was 81 % in the group of biopsied embryos cultured for lb and 98% in the group of biopsied embryos cultured for 15h, respectively. When biopsied embryos after ultrarapid freezing and thawing were transferred to the uteri of pseudopregnant recipients, normal live young were born. These results suggest that this freezing method can efficiently cryopreserve biopsied mouse embryos.

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MTT Assay Using Primary Cultured Rat Hepatocytes (백서의 초대 배양 간세포를 이용한 MTT assay$^{1)}$)

  • Ha, Hun;Yoon, Soo-Hong;Fujii, Takeru;Hori, Hitoshi
    • Environmental Analysis Health and Toxicology
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    • v.9 no.1_2
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    • pp.19-23
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    • 1994
  • The tetrazolium dye, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), is reduced by live but not dead cell, and this reaction is used as the end point in a rapid drug screening assay. It can also be used for accurate determinations of drug sensitivity but only if a quantative relationship is established between cell number and MTT-formazan production. Several conditions were examined to devise an in vitro assay method in primary cultured hepatocytes, such as optimum wavelength, optimal MTT concentration, optimal incubation time, and cell density.

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Morphology of one Frontonia and two Spathidium ciliates (Ciliophora: Intramacronucleata) from Korea

  • Park, Mi-Hyun;Min, Gi-Sik
    • Journal of Species Research
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    • v.6 no.spc
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    • pp.63-66
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    • 2017
  • Three ciliate species, Frontonia schaefferi Bullington, 1939, Spathidium curiosum Foissner, 2016, and S. muscicola Kahl, 1930, were collected from freshwater and terrestrial habitats in Korea. Their morphology was investigated based on live observations and protargol impregnation. Frontonia schaefferi was characterized by a cell size of $89-112{\times}45-53{\mu}m$ (after protargol impregnation) and three vestibular kineties. Spathidium curiosum was characterized by a cell size of ca. $125{\times}25{\mu}m$ in vivo and the unusual shape of the extrusomes. Spathidium muscicola was characterized by a cell size of ca. $135{\times}40{\mu}m$ in vivo and four to six macronuclear nodules. These three species are new records for Korea.

Development of Probiotic Products and Challenges (프로바이오틱 제품 개발 동향과 과제)

  • Seo, Jae-Gu;Lee, Gwa-Soo;Kim, Jin-Eung;Chung, Myung-Jun
    • KSBB Journal
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    • v.25 no.4
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    • pp.303-310
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    • 2010
  • Probiotics beneficially affect the health of the host via various mechanisms in the intestine. Recent developments in probiotic products have mainly been made to maximize probiotic effects in human. In this regard, probiotic products containing doubly coated or encapsulated cells, multi-species probiotics, or high viable cell number (1010 viable cells/gram or more) have been developed and are already available in the market. Until now, the majority of probiotics contain live cells but little attention has been paid to other alternative products such as heat-killed cell or bacteriocin-containing ones, which could have broad applications due to advantages over live cell-based probiotics, such as safety and stability. In addition, genetically engineered lactic acid bacteria could be of great importance in the field of alimentary health if they are carefully designed for biological safety. Although a number of probiotics are marketed by claiming health benefits, regulations for health claims will be more stringent. Therefore sufficient scientific and clinical evidences supporting the safety and efficacy of the potential probiotic strain will be required by the regulatory authority for a health claim, which thus may have a huge impact on the future probiotic market.

Cell-laden Gelatin Fiber Contained Calcium Phosphate Biomaterials as a Stem Cell Delivery Vehicle for Bone Repair (세포 함유 젤라틴 파이버 응용을 통한 골 재생 유도용 인산칼슘 생체재료 세포 탑재 연구)

  • Kim, Seon-Hwa;Hwang, Changmo;Park, Sang-Hyug
    • Journal of Biomedical Engineering Research
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    • v.43 no.1
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    • pp.61-70
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    • 2022
  • Natural and synthetic forms of calcium phosphate cement (CPC) have been widely used in bone repair and augmentation. The major challenge of injectable CPC is to deliver the cells without cell death in order to regenerate new bone. The study objective was to investigate for the potential of stem cell-laden gelatin fibers containing injectable, nanocrystalline CPC to function as a delivery system. Gelatin noddle fiber method was developed to delivered cells into nCPC. Experimental groups were prepared by mixing cells with nCPC, mixing cell-laden gelatin fibers with nCPC and mixing cell-laden gelatin fibers containing BMP-2 with nCPC. Media diffusion test was conducted after dissolving the gelatin fibers. SEM examined the generated channels and delivered cell morphology. Fibers mixed with nCPC showed physical setting and hardening within 20 min after injection and showed good shape maintenances. The gelatin fibers mixed nCPC group had several vacant channels generated from the dissolved gelatin. Particularly, proliferation and attachment of the cells were observed inside of the channels. While live cells were not observed in the cell mixed nCPC group, cells delivered with the gelatin fibers into the nCPC showed good viability and increased DNA content with culture. Cell-laden gelatin fiber was a novel method for cell delivery into nCPC without cell damages. Results also indicated the osteogenic differentiation of gelatin fiber delivered cells. We suggest that the cell-laden gelatin fibers mixed with nCPC can be used as an injectable cell delivery vehicle and the addition of BMP-2 to enhances osteogenesis.

Degradation of human immunoglobulins and cytotoxicity on HeLa cells by live Trichomonas vaginalis (질편모충 단백질분해효소의 세포독성 및 인체면역글로불린 분해능)

  • 민득영;류재숙
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.39-46
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    • 1997
  • The present study was undertaken to determine whether live T. uaginnlis degrades human secretory IgA, serum If and IgG molecules. Human immunoglobulins were exposed to live trophozoites, parasite Iysate, and excretory-secretory product (ESP) of T ucginnlis. To determine the fragmentation of immunoglobulins, the reaction sample was subjected to SDS-PAGE and EITB, and peroxidase conjugated antihuman IgA and IgG were used as probes. Live trophozoites degraded secretory IgA, serum IgA and IgG, and degradation were pressed forward by the prolongation of the incubation time and by increasing the number of trichomonads respectively. Also the Iysates and ESP of trichomonads degraded IgA and IgG. The cysteine and serine proteinase inhibitors such as I-64, antipain, iodoacetic acid, iodoacetamide, TLCK reduced the ability of cleaving immunoglobulins. The proteinase activity and cytotoxicity of T. uaginnlis to HeLa cells were decreased when live T. vusinalis was treated with metallo-proteinase inhibitor as well as cysteine and serine proteinase inhibitors. These results suggest that proteinase secreted from live T ucginclis may play a part role in host pathogenesis by T. uosinnlis, and the cleaving ability of host immunoglobulins by the proteinase may contribute as a one of immune evasion mechanism for parasite survival in the host.

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Evaluation of Six Species Ciliates as a Live Food and Culture Environment for Euplotes sp. (먹이생물로써의 섬모충 6종의 평가와 Euplotes sp.의 배양 환경)

  • Yoo Jin Hyung;Hur Sung Bum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.4
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    • pp.342-347
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    • 2002
  • Ciliates have the possibility of a new live food in marine finfish culture because of their wide range of body size, thin tell wall, show motility, and fast reproduction rate. In this research, six species of ciliates were isolated from south coast and salt pond in Korea. The fitness of these species as a live food was evaluated in terms of size, motility, suspensibility and cell density. As the result, Euplotes sp. (K-1) was found suitable to be a new live food which might substitute rotifers, Brachionus plintilis and B. rotundiformis in fish larvae culture. The modified $F{\emptyset}yn's$ Erdschreiberd media, MErds-2 with the addition of glycine, glucose and yeast extract increased six times higher growth rate of Euplotes sp. (K-1) than the basic F$\emptyset$yn's Erdschreiberd media. The optimum water temperature, pH and light intensity for this ciliates were $22.5^{\circ}C$, 8 and 2,000 lux, respectively, and its culture environmental range was relatively wide, On the other hand, this ciliate fed baker's yeast, Saccharomyces cererisiae grew up to 1,240 inds./mL with the inocula of 100 inds./mL within 7 days. The results of the study showed that Euplotes sp. (K-1) has a potential to be utilized as a new live food in fish larvae culture.