Results And Discussion

The expression of the ovalbumin gene is induced in accordance with the development of oviduct tissue, which in turn is developmentally regulated but is also possible to be induced artificially by administering steroid hormones to immature female chickens.6'7 We confirmed that laying hens and 12-day estradiol-treated chickens expressed ovalbumin either in the level of mRNA or of protein (data not shown). Untreated immature chickens did not express ovalbumin as expected. Oviducts from these...

References

1) Kool, M., Voncken, J.W., van Lier, F.L.J., Tramper, J. and Vlak, J.M., 1991, Detection and analysis of Autographa californica nuclear polyhedrosis virus mutants with defective interfering properties, Virology, Vol. 183, pp. 739-746. 2) Cusack, T. and Mccarth, W.J., 1989, Effect of serial passage on genetic homogeneity of plaque variant of Lymantria dispar nuclear polyhedrosis virus (Hamden LDP-67), Journal of General Virology, Vol. 70, pp. 2963-2972. 3) Lee, H.Y. and Krell, P.J., 1994,...

Results And Discussion 31 Three IgG1 variants

Three anti-CD20 antibodies were compared in this study standard IgG1 with fucose and native amino acids (STD-IgG1), defucosylated IgG1 with native amino acids (dFu-IgG1), and S298A E333A K334A- IgG1 with fucose (AAA-IgG1) (Figure 1). STD-IgG1 is equivalent to commercially available Rituxan, which is used in the treatment of non-Hodgkin lymphomas. Rituxan is produced by Chinese hamster ovary (CHO) cells. AAA-IgG1 was also produced by CHO cells whereas dFu-IgG1 was produced by rat hybridoma YB2 0...

Cotreatment of ERWPt suppresses twostage cell transformation foci

Few foci appeared in the control without MCA PMA treatment. In the presence of MCA PMA treatment, many large foci significantly appeared in the culture dishes. Appearance of transformation foci was strongly suppressed by treatment of ERW Pt (Table 1). Transformation foci were suppressed by addition of Pt nanoparticles to ERW in concentration-dependent manner. ERW supplemented with 1 ppm, 3 ppm and 10 ppm of Pt nanoparticles showed 57.1 , 65.4 and 100 of transformation foci suppression compared...

Thermodynamic And Kinetic Effects Of Human IgG1 Defucosylation On IgG1FCyRiiia Interaction

Akira Okazaki1, Emi Shoji-Hosaka1, Kazuyasu Nakamura1, Masako Wakitani1, Kazuhisa Uchida1, Shingo Kakita1, Kouhei Tsumoto2, Izumi Kumagai2 and Kenya Shitara1 1Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Aoba-yama 07, Aoba-ku, Sendai 980-8579, Japan Abstract Lack of fucose on human IgG1 oligosaccharide improves its affinity for Fcy...

HPLC analysis of Asnlinked oligosaccharides on antibodies

Antibody was purified with Protein A from culture medium and was dissolved in 100 mM sodium phosphate buffer (pH 7.2) containing 0.5 SDS and 50 mM 2-mercaptoethanol, and was then denatured at 100 C for 5 min. After dilution with 100 mM sodium phosphate buffer and the addition of 0.5 Nonidet P-40, 2.5 units of N-glycanase (Roche Diagnostics) was added to the sample, which was then incubated for 16 h at 37 C. Liberated Asn-linked oligosaccharides were separated from the salts and peptides using...

Assessment of cell survival and proliferation avoiding anoikis

The externalization of phosphatidylserine after the detachment of cells was examined by Annexin V binding. As shown in Fig. 1, 6M-4 cells were apparently anoikis resistant during the first 6h on both the MPC and polystylene matrix. A larger fraction of the parental MDCK cells on the MPC matrix presented double positive signals than those on the polystylene matrix. After a 6 camptothecin, which is an inhibitor of the gyrase responsible for DNA replication, the 6M-4cells were still resistant to...

Targeted Methods Analytical Platform

Phenomenome Discoveries Inc. is developing a Targeted Methods analytical platform for the investigation of major primary and secondary metabolites. Researchers can choose a combination of several preselected sets of primary and secondary metabolites in order to efficiently compare relevant biochemical levels in experimental samples. Our GC MS and LC MS analytical methods achieve excellent chromatographic separation of most metabolites, and by using SIM (single ion monitoring) and SRM (selected...

Grouping of 6M4 cells depending on growth in soft agar

As described above, 6M-4 cells consist of a morphologically heterogeneous population, sot, we attempted clonal selection using the dilution and soft agar culture methods. We found at least two populations in the 6M-4 cells, namely, one group capable of growing in soft agar and the other incapable of proliferating in soft agar but able to survive. Both populations maintained the characteristics of adherence to polystylene and resistance to anoikis and MEP. The former populations formed larger...

Materials Methods

Viral producer cells, GP293 Clontech , 293FT Invitrogen and GP293-G4 Mizuarai et al., 2001 cell lines were maintained in high glucose DMEM media Sigma supplemented with 10 FBS BioWest , 0.1 mM MEM non-essential amino acids Invitrogen , 2mM L-glutamine Sigma and penicillin-streptomycin Wako on collagen-coated dishes Iwaki . Virus producer cells were transfected with GFP or P-galactosidase lacZ coding virus vector plasmid and appropriate expression plasmids see Table 1 and Results to produce the...

Production Of Chimeric Antibodies By Transgenic Chicken Bioreactors

Yoshinori Kawabe, Akitsu Hotta, Ken-ichiro Ono, Kazuhisa Esaka, Ken-ichi Nishijima, Masamichi Kamihira and Shinji Iijima Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan Abstract Recombinant antibodies have been successfully used as pharmaceuticals for diseases such as cancer and rheumatoid, and an effective mass production method for therapeutic antibodies is urgently desired. Transgenic chicken proposed as a...

Introduction

Glucose has been used as a sugar source in most cell culture media. However, it is not always suitable for the production of biologicals because it generates a high concentration of lactic acid by glycolysis, leading to a decrease in medium pH below optimal range for cell and product stability. Many investigators have shown that fructose was better than glucose in slow utilization and low lactate production Imamura et al., 1982 Petch et al., 1996 . Despite these facts, fructose has been little...

Observation Of Individual Cell Behaviors To Analyze Mitogenic Effects Of Sericin

Tomohiro Toyosawa1, Satoshi Terada1, Masahiro Sasaki2, Hideyuki Yamada2, and Masahiro Kino-oka3 1Department of Applied Chemistry and Biotechnology, University of Fukui 3-9-1 Bunkyo, Fukui 910-8507, Japan 2Technology Department, Seiren Co., Ltd. 1-10-1 Keya, Fukui 9188560, Japan 3Department of Chemical Science and Engineering, Osaka University 1-3 Machikaneyama-cho, Toyonaka 560-8531, Japan Abstract Mammalian cell cultures are used in many fields. Various biologically active agents are produced...

Trace element optimized prototypes for other cell lines

A trace element optimized transfected PER.C6 medium, PER.C6 CD Prototype, supported cell growth to a higher maximum viable cell density as well as increased cell culture longevity in both small-scale Figure 3 and large-scale data not shown agitated suspension than unsupplemented and supplemented AEM and the commercially available control. The CD Prototype also supported higher antibody production than the control Figure 3 . Two HeLa formulations with optimized trace elements have been...

The intracellular H2O2scavenging effect of fucoidan on HT1080 cells

Then, we checked to determine whether fucoidan could act as an H2O2 scavenger. DCFH-DA has been used as a substrate for measuring intracellular H2O2 production in HT1080 cells. DCFH-DA is hydrolyzed by esterases to dichlorofluorescin DCFH , which is trapped within the cell. This nonfluorescent molecule is then oxidized to fluorescent dichlorofluorescin DCF by action of cellular H2O2. HT1080 cells were pre-treated with MEM containing fucoidan for 24h, then incubated with 5 M DCFH-DA for 30 min...

Neural stem cell cultivation

Neural stem cells NSCs were generated from E18 cortical tissue, processed and grown as neurospheres in non-agitated ultra low attachment ULA 6-well culture dish Corning in a 5 CO2, 37 C incubator 3-4 . The control medium was DMEM F12, 1x N2, 10 ng ml EGF and 20 ng ml basic FGF Invitrogen . The test medium was Neural sySTEM EX, a medium prototype, with 2 mM L-glutamine, 1x B27TM without retinol acetate RA , 10 ng ml EGF and 20 ng ml basic FGF. Viable cells were counted by trypan blue exclusion...

Springer

Catalogue record for this book is available from the Library of Congress. ISBN-13 978-1-4020-4457-1 e-book P.O. Box 17, 3300 AA Dordrecht, The Netherlands. All Rights Reserved 2006 Springer No part of this work may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording or otherwise, without written permission from the Publisher, with the exception of any material supplied specifically for...

Materials And Methods

We used normal human astrocyte Cambrex Bio Science, Walkersville, MD . NHA were seeded into 12-well plates for quantitative RT-PCR at a density of 2 x 104 well, or 24-well plates for MTT assay at a density of 1 x 104 well in ABM medium Cambrex Bio Science supplemented with 5 FCS, rhEGF and IGF, and cultured in a humidified atmosphere of 5 CO2 in 95 air at 37 C. Stock solutions of PLLA were made in dimethyl sulfoxide DMSO and final concentration of DMSO was 0.1 this concentration did not affect...

Trace Element Optimization Enhances Performance And Reproducibility Of Serumfree Medium

Kenerson1, Lia D. Tescione1, Dale F. Gruber1, David W. Jayme2, Donald G. Munroe1, and Stephen F. Gorfien1 1Invitrogen Corporation, 3175 Staley Road, Grand Island, New York 14072, USA Department of Biochemistry, Brigham Young University Hawaii, 55-220 Kulanui Street, Laie, Hawaii 96762, USA Abstract Optimizing the quantity and composition of trace elements in the AEM Adenovirus Expression Medium formulation, resulted in an improved version of AEM, which is a...

Results

We firstly studied effects of VSV-G on virus titer by changing the amount of the expression vector for the transfection, since the protein has cytotoxic effects on host cells Ory et al., 1996 . We found that 100 to 200 ng of VSV-G expression plasmid DNA per 24 well plate Iwaki gave a good result data not shown . We performed similar experiments on virus vectors and gag pol for 293FT cells expression plasmids to determine the amount of DNA which gave the best results data not shown . In the...

Using Metabolomics To Better Understand Functional Foods

Goodnowe1, Yasuyo Yamazaki1, Bernhard H. J. Juurlink2 1Phenomenome Discoveries Inc. 204-407 Downey Road, Saskatoon, SK, S7N 4L8, Canada 2Department of Anatomy and Cell Biology, College of Medicine, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada Abstract Decreasing hypertension and inflammation are important human health objectives. Functional foods are being developed to provide an alternative treatment for mild to moderate symptoms as well as for alternatives to...

Proteome Analysis Of Neurite Differentiation Of Phytoestrogentreated Pc12 Cells Preliminary results

Hiroko Isoda, Mariko Seki, Terence P. N. Talorete, Junkyu Han Graduate School of Life and Environmental Sciences, University of Tsukuba, Japan Abstract A novel isoflavone, 7, 2', 4'-trihydroxyisoflavone-4'-O-P- D-glucopyranoside was isolated from the aerial part of Crotalaria sessiliflora. The isoflavone glucoside was found to enhance the proliferation of the MCF-7 human breast cancer cell line, which possesses estrogen receptors ERs and responds in culture to estrogen. The estrogenic property...

Downregulation Of a5p1 Integrin Expression During Neuronal Differentiation In Neural Stem Cells

Dai Muramatsu, Naoko Yoshida, Sohei Hishiyama, Yusei Miyamoto, Tatsuhiro Hisatsune Department of integrated biosciences, University of Tokyo, Bioscience building 402, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan Abstract During neuronal differentiation, dynamic morphological and physiological changes found in neural stem cells imply the involvement of cell adhesion molecules. Integrins, which are major family of cell adhesion molecule play a central role in migration, proliferation and...

Addition of Trace Element Supplement to 293 II SFM

Stock cells were grown in 293 SFM II to a day 4 cell density of approximately 2.5 x 106 vc mL 99 viable and passaged into the four test conditions. Due to the relatively high density of the stock cells all passage 1 test cultures lagged. After two subsequent passages all conditions recovered and achieved day 4 cell densities of approximately 2.5 x 106 vc mL. These cultures were then passaged on day 6 after reaching cell densities in excess of 3.0 x 106 vc mL. The cells in the unsupplemented...

Cell culture on Toshi Substrata made of adult bovine pancreas

3.2.1 Effect of Section Thickness on Cell Morphology Pancreas-derived TOSHI-substrata cut into 5 or 10 .m-thick sections were prepared and the effect of section thickness on the culture was examined. HIT-T15 cells were seeded on the TOSHI-substrata and microscope observation revealed that they adhered and spontaneously aggregated on both sized sections Fig. 2 no significant difference was observed. This suggests that the TOSHI-substrata can induce cells to aggregate regardless of section...

Mesenchymal Cells Metalloproteinases And Adhesion On Microcarriers

Cytodex Beads

Mosbeux1, A.R. dos Santos Pedregal1, D. Ribeiro de Sousa1 , Vincianne Hendrick1, N. Joseph, M. Bensellam1, D. Blankaert1, T. Marique1, C. Alloin1, D. Parent2, C. Liesnard2, J.P. Van Vooren2, S. Lowagie1 and J. Werenne1 1Laboratory of Animal Cell Biotechnology. Dept. of Bioengineering, Faculty of Sciences, Universit Libre de Bruxelles, Belgium and 2Erasme Hospital,, Brussels, Belgium. Abstract The purpose of this work was to approach the development of a safe and validable process to grow the...