• Title/Summary/Keyword: Cell Observation

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Effects of high Cell Density on growth-Associated Monoclonal Antibody Production by Hybridoma T0405 Cells Immobilized in Macroporous Cellulose carriers

  • Hideki Mochoda;Wang, Pi-Chao;Fr Jr. Nayve;Ryuji Sato;Minoru Harige;Nakao Nomura;Masatoshi Matsumura
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.110-117
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    • 2000
  • Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.

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Application of Periodic Acid Thiocarbohydrazide Silver Proteinate Physical Development ( PA-TCH-SP-PD) Stain to Observation of Sertoli Cell (세르톨리세포 관찰을 위한 PA-TCH-SP-PD 염색의 적용)

  • 박영석;이성호
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.331-339
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    • 1998
  • The purpose of this study was to investigate the applicability of periodic acid thiocarbohy-drazide silver proteinate, physical development (PA-TCH-SP, -PD) stain to the seminiferous tubules for the ultrastructural studies of Sertoli cell column and Sertoli cell processes. In the Sertoli cell cloumn and Sertoli cell processes, high concentration of the reactive granules were observed under transmission electronmicroscope (TEM) after PA-TCH-SP-PD stain. Also some reactive granules were seen in the spermatogonium cytoplasm, clearly. These reactive granules specifically stained with PA-TCH-SP, -PD make the Sertoli cell column, Sertoli cell processes and spermatogonium cytoplasm easy to distinguish from nucleus of the germ cells, spermatocyte, spermatid and residual body which did not contain the reactive granules. This result indicates that the PA-TCH-SP, -PD stain is superior to other traditional electronic double stain methods for the ultrastructural studies of Sertoli cell.

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Mouse Embryo Culture used in Quality Control of Water for Human in Vitro Fertilization : The One-cell Stage Versus the Two-cell Stage Model (수질에 대한 1-세포기 및 2-세포기 생쥐배아를 이용한 생물학적 정도관리에 관한 연구)

  • Lee, Ye-Kyung;Chung, Hye-Won;Kim, Hyung-Mee;Oh, Seung-Eun;Son, Young-Soo;Yu, Han-Ki;Woo, Bock-Hee
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.1
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    • pp.9-17
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    • 1993
  • This study was carried out investigate the effect of water quality and the kind of media on the in vitro development of 1-cell and stage mouse embryos. $F_1$ hybrid mice were superovulated and timely mated. 1-cell stage and 2-cell stage mouse embryos were recruited and taken into Ham's F-10 or m-KRB media which was made of two of two kinds of water having different quality, highly purified water and tap water. 2-cell stage embryos grew up well in vitro to blastocyst or hatching blastocyst regardless of the composition of culture media, but 1-cell stage mouse embryo didn't develop well to blastocyst or hatching blastocyst in simple media like m-KRB. These results meant in vitro devleopment of 1-cell stage mouse embryo neded complex media like Ham's F-10 which contained abundant protein components. In case of quality control for water, in vitro fertilization program. observation of in vitro development of 2-cell mouse embryos up to blastocyst or hatching blastocyst media such as m-KRB would be efficatious in detecting the difference of water quality.

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Ginseng-derived nanoparticles induce skin cell proliferation and promote wound healing

  • Song Yang;Shuyan Lu;Limei Ren;Shuai Bian;Daqing Zhao;Meichen Liu;Jiawen Wang
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.133-143
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    • 2023
  • Background: Past studies suggested that ginseng extracts and ginseng-derived molecules exerted significant regulatory effects on skin. However, no reports have described the effects of ginseng-derived nanoparticles (GDNPs) on skin cell proliferation and wound healing. In this study, we investigated whether GDNPs regulate the proliferation of skin cells and promote wound healing in a mouse model. Methods: GDNPs were separated and purified via differential centrifugation and sucrose/D2O gradient ultracentrifugation. GDNP uptake, cell proliferation and cell cycle progression were measured by confocal microscopy, CCK-8 assay and flow cytometry, respectively. Cell migration and angiogenic effects were assessed by the wound scratch assay and tube formation assay, respectively. ELISA was used to detect extracellular matrix secretion. The relevant signaling pathway was confirmed by western blotting. The effects of GDNPs on skin wound healing were assessed by wound observation, HE staining, and western blotting. Results: GDNPs possessed the essential features of exosomes, and they were accumulated by skin cells. Treatment with GDNPs notably enhanced the proliferation of HaCaT, BJ and HUVECs. GDNPs also enhanced the migration in HaCaT cells and HUVECs and angiogenesis in HUVECs. GDNPs increased the secretion of MMP-1, fibronectin-1, elastin-1, and COL1A1 in all three cell lines. GDNPs regulated cell proliferation through the ERK and AKT/ mTOR pathways. Furthermore, GDNPs facilitated skin wound healing and decreased inflammation in a mouse skin wound model. Conclusion: GDNPs can promote skin wound healing through the ERK and AKT/mTOR pathways. GDNPs thus represent an alternative treatment for chronic skin wounds.

Development of Radar Polygon Method : Areal Rainfall Estimation Technique Based on the Probability of Similar Rainfall Occurrence (Radar Polygon 기법의 개발 : 유사강우발생 확률에 근거한 면적강우량 산정기법)

  • Cho, Woonki;Lee, Dongryul;Lee, Jaehyeon;Kim, Dongkyun
    • Journal of Korea Water Resources Association
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    • v.48 no.11
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    • pp.937-944
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    • 2015
  • This study proposed a novel technique, namely the Radar Polygon Method (RPM), for areal rainfall estimation based on radar precipitation data. The RPM algorithm has the following steps: 1. Determine a map of the similar rainfall occurrence of which each grid cell contains the binary information on whether the grid cell rainfall is similar to that of the observation gage; 2. Determine the similar rainfall probability map for each gage of which each grid cell contains the probability of having the rainfall similar to that of the observation gage; 3. Determine the governing territory of each gage by comparing the probability maps of the gages. RPM method was applied to the Anseong stream basin. Radar Polygons and Thiessen Polygons of the study area were similar to each other with the difference between the two being greater for the rain gage highly influenced by the orography. However, the weight factor between the two were similar with each other. The significance of this study is to pioneer a new application field of radar rainfall data that has been limited due to short observation period and low accuracy.

Cloning of the Genomic DNA Which Complements the Drug-Hypersensitivity of Saccharomyces cerevlsiae

  • Lee, Yun-Sik;Park, Kie-In
    • BMB Reports
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    • v.30 no.3
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    • pp.167-172
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    • 1997
  • The yeast Saccharomyces cerevisiae, mutant CH117, shows a drug-hypersensitivity (dhs) to cycloheximide, bleomycin, actinomycin D, 5-fluorouracil. nystatin, nigericin and several other antibiotics. CH 117 was also temperature-sensitive (ts). being unable to grow at $37^{\circ}C$ and secreted more invertase and acid phosphatase into the medium than the parent yeast. CH117 grows very slowly and the cell shape is somewhat larger and more sensitive to zymolyase than the wild type cells. Light microscopic and electron microscopic observation also revealed abnormality of the mutant cell wall. These characteristics indicate that CH117 has a defect in an essential component of the cell surface and that the cell wall which performs barrier functions has become leaky in the mutant. We screened a genomic library of wild type yeast for clones that can complement the mutation of CH117. A plasmid, pCHX1, with an insert of 3.6 kilobases (kbs) could complement the dhs and ts of CH117. Deletion and subcloning of the 3.6 kb insert showed that a gene for the complementation of mutant phenotypes was located in 1.9 kbs Puvll-Hindlll fragment.

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Ethanol Production by a New Method of Alginate-Immobilization (새로운 Alginate 고정화 방법에 의한 에탄올 생산)

  • Kim, Eun-Young;Kim, Seung-Wook;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.373-380
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    • 1993
  • When the cells of yeast K35 were immobilized in Ca-alginate gel, cell concentration and viability decreased as alginate concentration increased. Considering the results, 2% (w/v) Ca-alginate concentration would be suitable. Among various concentrations of additives and cross-lin-king agent, the addition of 1.67% (w/v) of bentonite together with 0.33% (v/v) of glutaraldehyde (ABG bead) resulted in the highest ethanol production of 1.8%(w/v), using YPD medium containing 2% glucose. ABG bead seemed to be more resistant to phosphate ion than Ca-alginate bead. 0.33%(w/v) of phosphate was a proper concentration for the ethanol production by ABG bead. Scanning electron microscopic observation depicted that the immobilized cells on the bead surface were coated by alginate gel and that the cells in the internal bead were cross-linked with alginate matrix. When repeated-batch culture was performed with ABG bead for 40 days in a packed-bed reactor, ethanol concentration of about 90~110 g/l-gel was maintained. Cell viability was maintained around 70%, and outgrowing cell concentration was below 6.3% of total cell concentration. Consequently, the results showed that ABG head was a potential carrier for continuous production of ethanol compared to conventional Ca-alginate bead.

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Scanning Electron Microscopic Observation on Early Vascular Lesion in Rat Lung Administered with Monocrotaline (Monocrotaline을 투여한 백서 폐의 초기 혈관병변에 관한 주사전자현미경적 관찰)

  • Park, In-Ae;Ham, Eui-Keun
    • Applied Microscopy
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    • v.21 no.1
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    • pp.86-107
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    • 1991
  • An experimental study was performed to observe the early effects of monocrotaline on pulmonary vascular system by means of light microscopy and scanning electron microscopy, attempting to expore the mechanism behind the process of pulmonary hypertension. Experimental animal(Sprague-Dawley male rats ; 150-200g B. W.) were intra-peritoneal administered with 100mg/kg B. W. monocrotaline. Authors observed light microscopically various gradational increase of wall thickness in pulmonary muscular and non-muscular arteries in duration from 2 weeks to 5 weeks after monocrotaline administration and the changes were more sever in the latter than the former. The scanning electron microscopy shows severe and diffuse endothelical cell swelling, microvilli and microbleb formation since 1 hour after monocrotaline administration and during the course, after 5 hours the severity of endothelial cell damage was prominent with presence of fibrin, webs, platelet thrombi and white cell adherence. It was concluded that the monocrotaline primarily induced severe and diffuse endothelial cell damage of pulmonary arteries and laterly added the participation of platelets, which attributed to the pathogenesis of monocrotaline induced pulmonary vascular lesions in relation to pulmonary hypertension.

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The Differentiation of the Female Gonial Cell in Echiuroid (Urechis unicinctus): A Fine Structural Study (개불(Urechis unicinctus) 자성생식세포의 분화과정에 관한 미세구조연구)

  • Choe, Rim-Soon;Shin, Kil-Sang;Joo, Chung-No;Hwang, Dae-Yeon
    • Applied Microscopy
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    • v.18 no.1
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    • pp.77-91
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    • 1988
  • Since the Urechis unicinctus-oocyte grows asynchronously in the body fluid, various oocytes in developmental stages can be prepared from each individual. The oocytes obtained from the coelomic fluid are then classified into five developmental stages according to the fine structural features. The earlier oocytes (${\sim}18{\mu}m$) form cluster and thereafter the oocytes grow singly without a distinct support of somatic cell, such as accessory cell or matrix cell. The yolk granules begin to appear already in the oocyte of cluster stage, however, the typical yolk was observed at the stage IV. Therefore, it was suggested that the yolk deposition is correlated with the coelomic fluid. The mature oocyte measured about $150{\mu}m$ produces the invagination not only on oolemma(indentation) but also on nuclear envelope. After the formation of the indentation, the mature ooytes are stored in storge sacs. The fine structural features were combined in aspect of structural concept of light microscopical observation.

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Immunohistochemical and Pathological Findings im Mice Inoculated with Encephalomyocarditis Virus (뇌심근염 바이러스 접종 마우스에 대한 병리학적 및 면역조직화학적 소견)

  • 신창호;조성환
    • Korean Journal of Veterinary Service
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    • v.17 no.1
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    • pp.67-80
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    • 1994
  • 8-10 week old ICR mice were infected intracerebrally and intraperitoneally with different encephalomyocarditis virus(K$_3$, $K_{11}$, ATT-VR 129) to observe histopathological and immunohistochemical change. Results obtained throuh the experiments were summarized as follows : 1. No differences in clinical signs by the virus strains and the inoculation routes were found. Mice infected with EMCV showed clinical signs after 3 days of inoculation. Main clinical signs were tremors, convulsions, circling movement, and uni or bilateral hindleg paralysis followed by death on the 3-8 days. In general, most of the infected animals died or recovered closely on the 8th day of postinoculation. 2. At necropsy, petechial and ecchymotic hemorrhages in lung were observed and no specific findings in other were observed. 3. In histopathological observation, neuroal cell degeneration perivascular mononucear cell in-filtration gliosis were appeared in central nervous system. Myocarditis with myocardial degeneration and necrosis, calcification were observed along with acinar cell necrosis of exocrine glands in pancreas, severe glomerulonephritis in kidney. Also, focal necrosis of hepatocytes and interstitial pneymonia hyperemia, hemorrhages in lungs were observed. 4. By immunohistochemical staining using ABCIT method, the positive cells were recognized in intracytoplasm of acinar cell in pancreas and intracytoplasm of neuronal cells in cerebrum.

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