Rapid titration of adenoviral infectivity by flow cytometry in batch culture of infected HEK293 cells.


:There is a constant and growing interest in exploitingadenoviruses as vectors for gene therapy when transientexpression of a therapeutic protein is necessary. Therequirement for an increased viral titre has prompted asearch for techniques by which this virus may be assayedwith greater speed and simplicity. Conventional plaqueassay for quantification of adenoviral vectors titre incurrent use is laborious and time-consuming (up to 14days). We report herein a method for the monitoring ofadenovirus expressing green fluorescent protein thatincorporates rapid and easy sample handling by means offlow cytometric analysis. Cells (HEK293) were infectedwith adenovirus at various multiplicity of infection(MOI), harvested 17 to 20 h post infection and analysedby flow cytometry. Assumptions were made that onefluorescent cell was infected by a single infectiousparticle at a relatively low MOI. The adenoviral titrewas subsequently estimated from cell analysis in arelatively short time. The results obtained with an E1-complementing cell line (HEK293) were compared with thatobtained using a non-complementing cell line (A549). APoisson distribution successfully modelled the profile ofinfection as a function of MOI. This provided a betterunderstanding of adenoviral infection at the earlieststage possible. Monitoring of GFP fluorescence and viruspropagation in a batch culture of infected cells wassubsequently used as a practical application of thevalidated method.






Gueret V,Negrete-Virgen JA,Lyddiatt A,Al-Rubeai M




Has Abstract


2002-01-01 00:00:00












  • Evaluating osteochondral defect repair potential of autologous rabbit bone marrow cells on type II collagen scaffold.

    abstract::The feasibility of using genipin cross-linked type II collagen scaffold with rabbit bone marrow mesenchymal stem cells (RBMSCs) to repair cartilage defect was herein studied. Induction of RBMSCs into chondrocytic phenotype on type II collagen scaffold in vitro was conducted using TGF-β 3 containing medium. After 3-wee...


    pub_type: 杂志文章


    authors: Chen WC,Yao CL,Wei YH,Chu IM

    更新日期:2011-01-01 00:00:00

  • Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays.

    abstract::This study was undertaken to evaluate genotoxic potential of Thermopsis turcica aqueous extracts on the roots of onion bulb (Allium cepa L.) by comet assay and random amplified polymorphic DNA technique. The Allium root growth inhibition test indicated that the EC50 and 2×EC50 values were 8 and 16 mg/ml concentrations...


    pub_type: 杂志文章


    authors: Ciğerci İH,Cenkci S,Kargıoğlu M,Konuk M

    更新日期:2016-08-01 00:00:00

  • DESI-MS as a tool for direct lipid analysis in cultured cells.

    abstract::Desorption electrospray ionization may be used as a fast and convenient method for analysis and identification of lipids in the cell culture. Oxidative stress, which usually involves changes in lipids, was used as a model of pathology to show the utility of this analysis methodology. This paper addresses the surface p...


    pub_type: 杂志文章


    authors: Bodzon-Kulakowska A,Cichon T,Golec A,Drabik A,Ner J,Suder P

    更新日期:2015-12-01 00:00:00

  • The enhancement of growth and differentiation of rat adrenal nerve cells by the addition of conditioned medium from human fibroblast cultures.

    abstract::The growth of rat adrenal nerve cells was remarkably enhanced by supplementing the cultured medium from the human fibroblast cell line, Hs 68. Maximum specific growth rate and length of the neurites were observed as 0.076 (1/hr) and 0.026 mm, respectively in 20% supplement of five day old medium. In adding more than 2...


    pub_type: 杂志文章


    authors: Hong JS,Kim DS,Kim SH,Choi DH,Lee JH,Lee HY

    更新日期:1998-03-01 00:00:00

  • A continuous multistage roller reactor for animal cell culture: 1. Patterns of growth, production and catabolism of a murine hybridoma.

    abstract::A Tubular Liquid Film Reactor was designed as a model system to transfer a batch culture kinetic to a continuous cascade. Cell density, product formation and substrate consumption rates were followed during fermentation at two dilution rates. In spite of the high dilution rates effective in each segment by itself high...


    pub_type: 杂志文章


    authors: Borth N,Steindl F,Weigang F,Reiter M,Katinger H

    更新日期:1990-05-01 00:00:00

  • Mutagenic and cytotoxic activities of benfuracarb insecticide.

    abstract::Benfuracarb is a carbamate insecticide used to control insect pests in vegetables and it has anti-acetylcholinesterase activity lower than other carbamates. Cytotoxic effects of benfuracarb were evaluated by using root growth inhibition (EC50), mitotic index (MI), and mitotic phase determinations on the root meristem ...


    pub_type: 杂志文章


    authors: Eren Y,Erdoğmuş SF,Akyıl D,Özkara A

    更新日期:2016-08-01 00:00:00

  • Silencing of soluble epoxide hydrolase 2 gene reduces H2O2-induced oxidative damage in rat intestinal epithelial IEC-6 cells via activating PI3K/Akt/GSK3β signaling pathway.

    abstract::Oxidative stress plays a vital role in the occurrence and development of intestinal injury. Soluble epoxide hydrolase 2 gene (EPHX2) is a class of hydrolytic enzymes. We aim to explore the effects and molecular mechanism of siEPHX2 on H2O2-induced oxidative damage in rat intestinal epithelial IEC-6 cells. IEC-6 cells ...


    pub_type: 杂志文章


    authors: Li J,Luo J,Zhang Y,Tang C,Wang J,Chen C

    更新日期:2020-02-01 00:00:00

  • A modified preplate technique for efficient isolation and proliferation of mice muscle-derived stem cells.

    abstract::We modified an existing protocol to develop a more efficient method to acquire and culture muscle-derived stem cells (MDSCs) and compared the characteristics of cells obtained from the two methods. This method is based on currently used multistep enzymatic digestion and preplate technique. During the replating process...


    pub_type: 杂志文章


    authors: Xu Z,Yu L,Lu H,Feng W,Chen L,Zhou J,Yang X,Qi Z

    更新日期:2018-12-01 00:00:00

  • Apoptosis resistance in tumor cells.

    abstract::Various antitumor agents induce apoptotic cell death in tumor cells. Since the apoptosis program in tumor cells plays a critical role in the chemotherapy-induced tumor cell killing, it is suggested that the defect in the signaling pathway of apoptosis could cause a new form of multidrug resistance in tumor cells. This...


    pub_type: 杂志文章


    authors: Mashima T,Seimiya H,Chen Z,Kataoka S,Tsuruo T

    更新日期:1998-09-01 00:00:00

  • Anti-allergic triterpenes isolated from olive milled waste.

    abstract::The degranulation of basophils stimulated by antigen is one of the allergic mechanisms of immediate hypersensitivity reactions. In a previous study, the β-hexosaminidase release of rat basophilic leukemia (RBL-2H3) cells stimulated by IgE cross-linking was specifically inhibited by ethanol extract of olive milled wast...


    pub_type: 杂志文章


    authors: Kishikawa A,Amen Y,Shimizu K

    更新日期:2017-04-01 00:00:00

  • The organotypic culture of human skin keratinocytes and fibroblasts to achieve form and function.

    abstract::We describe an organotypic model of human skin comprised of a stratified layer of human epidermal keratinocytes and dermal fibroblasts within a contracted collagen lattice. Feasible and reproducible production of the skin construct has required the use of traditional as well as specialized culture techniques. The conf...


    pub_type: 杂志文章


    authors: Parenteau NL,Bilbo P,Nolte CJ,Mason VS,Rosenberg M

    更新日期:1992-01-01 00:00:00

  • Inhibitory effects of chlorogenic acid and its related compounds on the invasion of hepatoma cells in culture.

    abstract::Actions of chlorogenic acid, a major component of coffee, andits constituents, caffeic and quinic acids, on theproliferation and invasion of AH109A, a rat ascites hepatomacell line, were investigated using in vitro assay systems. Allthree components suppressed the AH109A invasion atconcentrations of 5-40 muM without a...


    pub_type: 杂志文章


    authors: Yagasaki K,Miura Y,Okauchi R,Furuse T

    更新日期:2000-07-01 00:00:00

  • Structure-activity relationship of antioxidants for inhibitors of linoleic acid hydroperoxide-induced toxicity in cultured human umbilical vein endothelial cells.

    abstract::Structure-activity relationship of antioxidants for the protective effects on linoleic acid hydroperoxide (LOOH)-induced toxicity were examined in cultured human umbilical vein endothelial cells. alpha-Tocopherol, 2,2,5,7,8-pentamethylchroman-6-ol, butylated hydroxytoluene, probucol, and fatty acid esters of ascorbic ...


    pub_type: 杂志文章


    authors: Kaneko T,Baba N,Matsuo M

    更新日期:2001-01-01 00:00:00

  • MiR-206 inhibits epilepsy and seizure-induced brain injury by targeting CCL2.

    abstract::To determine the function of miR-206 in epilepsy. Epileptic rat model was established by intra-amygdala injection of kainic acid (KA). Expression levels of miR-206, C-C Motif Chemokine Ligand 2 (CCL2) and interleukin-1β (Il-1β) in hippocampus tissues was measured by reverse transcription-quantitative PCR (RT-qPCR) and...


    pub_type: 杂志文章


    authors: Wu Z,Liu Y,Huang J,Huang Y,Fan L

    更新日期:2019-06-26 00:00:00

  • Evaluation of the antiviral effect of new compounds by using cellular cultures.

    abstract::Replication Kinetics of HIV was studied under single-growth cycle conditions in C8166 lymphoblastoid cells by back-titrating released and cell-associated infectious virus et each time point. Under these conditions HIV seemed to replicate faster than previously estimated. The amount cell-associated virus always exceede...


    pub_type: 杂志文章


    authors: Dianzani F,Antonelli G

    更新日期:1991-02-01 00:00:00

  • A direct computer control concept for mammalian cell fermentation processes.

    abstract::In the last 10 years, new assignments and the special demands of mammalian cells to the culture conditions caused the development of complex small scale fermentation setups. The use of continuous fermentation and cell retention devices requires appropriate process control systems. An arrangement for control and data-a...


    pub_type: 杂志文章


    authors: Büntemeyer H,Marzahl R,Lehmann J

    更新日期:1994-01-01 00:00:00

  • Electroporation: an arsenal of application.

    abstract::Electroporation is a way to induce nanometersized membrane pore for exogenous substances delivery into cytoplasm using an artificial electric field. Now it was widely used for molecules transfer especially in molecular experiments and genetic aspects. In recent years, modern electroporation on the embryo was developed...


    pub_type: 杂志文章


    authors: Yuan TF

    更新日期:2007-06-01 00:00:00

  • Use of cell cultures, nuclear and radioanalytical techniques for metallotoxicological studies at the JRC-Ispra.

    abstract::This paper reviews the JRC-Ispra research activity on toxicological studies related to the environmental and occupational exposure of trace metals, as carried out by a combination of cell culture methodologies with nuclear and radioanalytical techniques.Applications concern the setting of uptake-effect relationships a...


    pub_type: 杂志文章


    authors: Sabbioni E,Pozzi G,Fischbach M,Bahbouth E

    更新日期:1993-01-01 00:00:00

  • Relative survivability of human osteoblasts is enhanced by 39 °C and ascorbic acid after exposure to photopolymerization ingredients.

    abstract::Photopolymerizable hydrogels offer great potential in cartilage tissue engineering due to their ability to conform to irregular defect shapes and be applied in a potentially minimally invasive manner. An important process requirement in the use of photopolymerizable hydrogels is the ability of the suspended cells to w...


    pub_type: 杂志文章


    authors: Dua R,Ramaswamy S

    更新日期:2013-08-01 00:00:00

  • A luciferase complementation assay system using transferable mouse artificial chromosomes to monitor protein-protein interactions mediated by G protein-coupled receptors.

    abstract::G protein-coupled receptors (GPCRs) are seven-transmembrane domain receptors that interact with the β-arrestin family, particularly β-arrestin 1 (ARRB1). GPCRs interact with 33% of small molecule drugs. Ligand screening is promising for drug discovery concerning GPCR-related diseases. Luciferase complementation assay ...


    pub_type: 杂志文章


    authors: Uno N,Fujimoto T,Komoto S,Kurosawa G,Sawa M,Suzuki T,Kazuki Y,Oshimura M

    更新日期:2018-12-01 00:00:00

  • Differences in developmental competence and gene expression profiles between buffalo (Bubalus bubalis) preimplantation embryos cultured in three different embryo culture media.

    abstract::The objective of this study was to compare effects of in vitro culture systems on embryonic development and expression patterns of developmentally important genes in preimplantation buffalo embryos. After IVM/IVF presumptive zygotes were cultured in one of three systems: undefined TCM-199, mCR2aa medium supplemented w...


    pub_type: 杂志文章


    authors: Sadeesh EM,Selokar NL,Balhara AK,Yadav PS

    更新日期:2016-10-01 00:00:00

  • Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model.

    abstract::In construction of epithelial cells as multilayers, the cells are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted to air to promote their stratification. We recently demonstrated that gingival epithelial cells form uniform monolayers on semi-permeable nitrocellulose membranes, supp...


    pub_type: 杂志文章


    authors: Gursoy UK,Gursoy M,Könönen E,Sintim HO,Uitto VJ,Syrjänen S

    更新日期:2016-12-01 00:00:00

  • Role of a putative tyrosine-O-sulfate receptor in the targeting and/or intracellular transport of tyrosine-sulfated proteins.

    abstract::By employing the affinity gel fraction technique, we have detected a 175 kDa tyrosine-O-sulfate (TyrS)-binding protein in sodium choleate extracts of the microsomal membrane fractions of bovine liver and pancreas, as well as canine liver and pancreas. Western blot analysis revealed the presence of the bovine liver Tyr...


    pub_type: 杂志文章


    authors: Liu MC,Sakakibara Y,Suiko M

    更新日期:1997-01-01 00:00:00

  • Automated time-lapse microscopy and high-resolution tracking of cell migration.

    abstract::We describe a novel fully automated high-throughput time-lapse microscopy system and evaluate its performance for precisely tracking the motility of several glioma and osteoblastic cell lines. Use of this system revealed cell motility behavior not discernable with conventional techniques by collecting data (1) from cl...


    pub_type: 杂志文章


    authors: Fotos JS,Patel VP,Karin NJ,Temburni MK,Koh JT,Galileo DS

    更新日期:2006-05-01 00:00:00

  • Identification of the pivotal role of glutamate in enhancing insect cell growth using factor analysis.

    abstract::Although the insect cell/baculovirus system is an important expression platform for recombinant protein production, our understanding of insect cell metabolism with respect to enhancing cell growth capability and productivity is still limited. Moreover, different host insect cell lines may have different growth charac...


    pub_type: 杂志文章


    authors: Chou CC

    更新日期:2014-10-01 00:00:00

  • Insights into protective effects of medium additives on animal cells under fluid stresses: the hydrophobic interactions.

    abstract::Animal cells in suspension culture can suffer severe mechanical damage from bursting gas bubbles or other hydrodynamic force sources. Certain chemical additives in the culture media, particularly some surface-active chemicals, can effectively protect animal cells against such damage. Previously we proposed that the pr...


    pub_type: 杂志文章


    authors: Wu J

    更新日期:1996-01-01 00:00:00

  • Insect cells respiratory activity in bioreactor.

    abstract::Specific respiration rate ( [Formula: see text]) is a key parameter to understand cell metabolism and physiological state, providing useful information for process supervision and control. In this work, we cultivated different insect cells in a very controlled environment, being able to measure [Formula: see text]. Sp...


    pub_type: 杂志文章


    authors: Pamboukian MM,Jorge SA,Santos MG,Yokomizo AY,Pereira CA,Tonso A

    更新日期:2008-05-01 00:00:00

  • Intracellular localisation studies of doxorubicin and Victoria Blue BO in EMT6-S and EMT6-R cells using confocal microscopy.

    abstract::The subcellular localisation of doxorubicin and Victoria Blue BO (VBBO) in a murine mammary tumour cell line EMT6-S, and the resistant sub-lineEMT6-R was studied, using confocal microscopy, in order to investigate their sites of action. In cells treated with doxorubicin (10 mu M) for 90 min, the pattern of intracellul...


    pub_type: 杂志文章


    authors: Burrow SM,Phoenix DA,Wainwright M,Tobin MJ

    更新日期:2002-05-01 00:00:00

  • Early detection of cell metabolism with a silicon microsensor.

    abstract::A silicon microsensor (ISFET - Ion Sensitive Field Effect Transistor) has been used to detect the metabolism of a cell population cultured on a coverslip and positioned close to the sensor surface. The system output is analyzed as a function of cell density. ...


    pub_type: 杂志文章


    authors: Martinoia S,Meloni M,Parodi MT,Tedesco M,Ciccarelli C,Grattarola M

    更新日期:1993-01-01 00:00:00

  • PLCgamma2 Activates CREB-dependent Transcription in PC12 Cells Through Phosphorylation of CREB at Serine 133.

    abstract::The cAMP and Ca(2+) signaling pathways activate the transcription factor CREB through its phosphorylation at Serine 133. Activation of CREB is involved in the regulation of various biological phenomena. To understand further the mechanisms of the regulation of CREB activity in response to activation of the cAMP and Ca...


    pub_type: 杂志文章


    authors: Iwamoto T,Mamiya N,Masushige S,Kida S

    更新日期:2005-01-01 00:00:00