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  • Articles  (11,783)
  • Chemical Engineering  (11,634)
  • cell culture  (77)
  • biofilm  (72)
  • Organic Chemistry
  • Process Engineering, Biotechnology, Nutrition Technology  (11,783)
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  • Articles  (11,783)
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  • 1
    Electronic Resource
    Electronic Resource
    Pilot production of u-PA with porous microcarrier cell culture (2000)
    Springer
    Cytotechnology 33 (2000), S. 13-19 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; porous microcarrier ; prourokinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A recombinant DNA CHO cell line secretingurokinase-type plasminogen activator (u-PA) wascultivated with Cytopore cellulose porousmicrocarriers in a 30l Biostat UC stirred tankreactor. After 26 days of culture, using a spinfilter toretain cells in bioreactor, the cell density couldreach 1.33 × 107 ml-1. The maximal u-PAactivity in supernatant was 7335 IU·ml-1, and204l supernatant containing 7.1 g u-PA was harvested.After 100 days of culture with 0.1% fetal bovineserum medium, a modified cell retention system whichcan be washed-out backward, substituted thespinfilter to prevent filter clogging. The maximalcell density was over 107 ml-1, the maximalu-PA activity in supernatant reached 6250IU·ml-1, and 1604l supernatant containing about51 g u-PA was harvested. Compared to perfusionculture, batch medium-replaced culture could raiseutilizing efficiency of the medium, increase cell specificproductivity and improve the quality of the product which wasnot steady in a 37 °C environment. Cells can movefrom seed porous microcarriers occupied by cells tovacant microcarriers spontaneously, withouttrypsinization, and continue to grow until all microcarriers contained cells. It shows that Cytoporeporous microcarriers are very useful and convenient toscale up cultivation step by step.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008127310890
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  • 2
    Electronic Resource
    Electronic Resource
    Flow Injection Analysis for Simultaneous Quantification of Prolactin Concentration and Glycosylation Macroheterogeneity in Cell Culture Samples (2000)
    Springer
    Cytotechnology 34 (2000), S. 237-242 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; flow injection analysis ; glycosylation ; macro-heterogeneity ; prolactin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A flow injection analysis (FIA) system is presented for a twostep immunoassay-based determination of the total humanprolactin (hPRL) concentration along with its degree ofglycosylation. Separate measurement of total hPRL and nonglysosylated human prolactin (nG-hPRL) were made using twoflow-through cartridges each containing immobilized antibodiesof different specificity. The antibodies are immobilized on thesurface of a carrier. Glycosylated hPRL (G-hPRL) and, thus, thedegree of glycosylation were calculated by the differencebetween the two specific determinations. Enhanced specificityfor the determination of nG-hPRL was obtained using unfavorablebinding conditions through incorporation of alkaline pH andchaotropic agents into the carrier/dispersion buffer. The assayfor total hPRL and nG-hPRL were each found to be linear withinthe relevant concentration range. The results of the two-stepFIA method were found to agree with those obtained by thestandard methods of ELISA and western blotting while offeringthe advantage of minimal analysis time (10 min) and eliminationof manual manipulations.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008170023099
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  • 3
    Electronic Resource
    Electronic Resource
    Production of interferon-β in a culture of fibroblast cells on some polymeric films (2000)
    Springer
    Cytotechnology 34 (2000), S. 165-173 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; enhanced production ; fibroblast cells ; interferon-β ; Langmuir-Blodgett film
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Normal human skin (NB1-RGB) cells were cultured in the presenceof polyinosinic and polycytidylic acids, diethylaminoethyldextran, cycloheximide and actinomycin D, which induced humaninterferon-β. The simplest induction method, that requiredonly polyinosinic and polycytidylic acids and diethylaminoethyldextran was found to give the highest production ofinterferon-β by the cells. The cell growth and productionof interferon-β were investigated for NB1-RGB cellscultured on silk fibroin, poly(γ-methyl-L-glutamate),poly(γ-benzyl-L-glutamate) and collagen films prepared bythe Langmuir-Blodgett (LB) and casting methods. The cell densityof NB1-RGB cells cultured on the LB films was found to be higherthan that on the cast films made of the same polymer. Thisindicates that not only the chemical structure of the polymersused for the preparation of the films but the preparationmethods of the films, i.e., casting and LB methods, are also astrong factor affecting the cell growth. The production ofinterferon-β per unit number of cells was found to behigher on the cast films than that on the LB films made of thesame polymer. This is explained by the fact that the optimalsuppressed growth of NB1-RGB cells on the cast films leads tothe enhanced production of interferon-β on the cast filmscompared to those on the LB films prepared by the same polymer.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008130223190
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  • 4
    Electronic Resource
    Electronic Resource
    Growing colorectal tumors: minimizing microbial and stromal competition and assessing in vitro selection pressures (2000)
    Springer
    Cytotechnology 34 (2000), S. 205-211 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; colorectal cancer ; microbial contamination ; stroma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Development of primary colorectal cancer cell lines ishampered by contamination from regional microbes, overgrowthof stromal cells, and purported genetic drift from selectionpressures in vitro. We initiated 32 primaryadenocarcinomas, 3 recurrences and 6 distant metastases incell culture. Twelve cell lines from eleven tumors weregenerated (26.8%) overall. Nine of 32 primary tumorsyielded 10 cell lines, 5 were lost to contamination, 13 wereoverwhelmed by stromal cells, and 5 demonstrated no growth.Addition of isobutyl methyl xanthine (IBMX) to culturelimited fibroblastoid growth. There was no associationbetween tumor location (p = 0.535, mid-P), degree ofdifferentiation (p = 0.850, mid-P) or clinicopathologic stage(p = 0.400, mid-P), and the ability of cells to becomeestablished in culture. The majority of cell lines hadsimilar nuclear DNA content and expression of cell-surfaceantigens compared with their parent tumors. Microbialcontamination and stromal cell overgrowth present thegreatest obstacle to capturing a representative bank ofcolon tumors in vitro.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008131428992
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  • 5
    Electronic Resource
    Electronic Resource
    Comparison of cell growth in T-flasks, in micro hollow fiber bioreactors, and in an industrial scale hollow fiber bioreactor system (2000)
    Springer
    Cytotechnology 34 (2000), S. 111-119 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; hollow fiber bioreactor ; hybridoma ; micro bioreactor ; optimization ; T-flask
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In this article, cell growth in a novel micro hollow fiberbioreactor was compared to that in a T-flask and theAcuSyst-Maximizer®, a large scale industrial hollowfiber bioreactor system. In T-flasks, there was relativelylittle difference in the growth rates of one murine hybridomacultured in three different media and for three other murinehybridomas cultured in one medium. However, substantialdifferences were seen in the growth rates of cells in themicro bioreactor under these same conditions. These differencecorrelated well with the corresponding rates of initial cellexpansion in the Maximizer. Quantitative prediction of thesteady-state antibody production rate in the Maximizer was moreproblematic. However, conditions which lead to faster initialcell growth and higher viable cell densities in the microbioreactor correlated with better performance of a cell line inthe Maximizer. These results demonstrate that the microbioreactor is more useful than a T-flask for determining optimalconditions for cell growth in a large scale hollow fiberbioreactor system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008167713696
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  • 6
    Electronic Resource
    Electronic Resource
    Justification of continuous packed-bed reactor for retroviral vector production from amphotropic ΨCRIP murine producer cell (2000)
    Springer
    Cytotechnology 34 (2000), S. 151-158 
    Details
    ISSN: 1573-0778
    Keywords: anchorage-dependent cell ; cell culture ; packed-bedreactor ; retroviral vector ; viral production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract To indentify a plausible large-scale production system forretroviral vector, three culture systems, i.e., batch culturewith medium exchange, microcarrier culture, and packed-bedreactor culture were compared. In batch cultures with mediumexchange, high cell concentrations were maintained for about amonth, and the harvested retroviral titer remained constant. Inmicrocarrier cultures, although cell growth was rapid, theretroviral titer was unexpectedly low, suggesting that the lowtiter was due either to serious damage to the retroviral vectoror to a reduction in the production rate of retroviral vector,caused by mechanical shear forces. Although the retroviral titer(maximum titer, 1.56 × 106) in the packed-bedreactor was a little bit lower than that obtained in the batchculture with medium exchange (maximum titer, 1.91 ×106), continuous production made it possible to increasethe cumulative titer up to 16-fold of that from the batchculture with medium exchange. Moreover, as the packed-bedreactor system requires less labor and shows excellentvolumetric productivity in comparison to batch cultures withmedium exchanges, it will be an appropriate production systemfor retroviral vector in large quantities.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008120313175
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  • 7
    Electronic Resource
    Electronic Resource
    The changing role of cell culture in the generation of transgenic livestock (1999)
    Springer
    Cytotechnology 31 (1999), S. 3-8 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; livestock ; milk ; nuclear transfer ; transgenic
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Transgenesis may allow the generation of farm animals with altered phenotype, animal models for research and animal bioreactors. Although such animals have been produced, the time and expense involved in generating transgenic livestock and then evaluating the transgene expression pattern is very restrictive. If questions about the ability and efficiency of expression could be asked solely in vitro rapid progress could be achieved. Unfortunately, experiments addressing transcriptional control in vitro have proved unreliable in their ability to indicate whether a transgene will be transcribed or not. However, initial studies suggest that cell culture may be able to predict in vivo post-transcriptional events. We review these issues and propose that strategies which engineer the transgene integration site could enhance the probability for efficient expression. This approach has now become feasible with the development of techniques allowing animals to be generated from somatic cells by nuclear transfer. The important step in this procedure is the use of cells grown in culture as the source of genetic information, allowing the selection of specific transgene integration events. This technology which has dramatically increased the potential use of transgenic livestock for both agricultural and biotechnological applications, is based on standard cell culture methodology. We are now at the start of a new era in large animal transgenics.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008044517150
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  • 8
    Electronic Resource
    Electronic Resource
    A simple and rapid reverse transcriptase assay for the detection of retroviruses in cell cultures (1999)
    Springer
    Cytotechnology 29 (1999), S. 221-227 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; polymerase chain reaction ; retrovirus ; reverse transcriptase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Reverse transcriptase (RT) is a good diagnostic tool for the detection of retroviruses. We have developed a simple and rapid assay for RT activity in culture supernatants. A 370-base RNA sequence from the tetracycline-resistance gene in pBR322 plasmid DNA was used as a template for RT-mediated cDNA synthesis. To detect the resultant cDNA, we used the nested polymerase chain reaction. A sensitivity test using purified recombinant RT of human immunodeficiency virus type 1 demonstrated that the detection limit of this method was 10-7–10-8 units of RT activity in 20 μl of a test sample (2 × 10-9–2 × 10-10 units ml-1). This method detected RT activity in unconcentrated supernatants of cell cultures infected with human T-cell leukemia virus, Moloney murine leukemia virus, Moloney murine sarcoma virus, or Rous sarcoma virus. This nonisotopic method provides results within 10 h and is useful for quality control to detect retroviruses in cell cultures.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008000210125
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  • 9
    Electronic Resource
    Electronic Resource
    Disposable bioreactor for cell culture using wave-induced agitation (1999)
    Springer
    Cytotechnology 30 (1999), S. 149-158 
    Details
    ISSN: 1573-0778
    Keywords: bioreactor ; cell culture ; disposable ; wave agitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract This work describes a novel bioreactor system for the cultivation of animal, insect, and plant cells using wave agitation induced by a rocking motion. This agitation system provides good nutrient distribution, off-bottom suspension, and excellent oxygen transfer without damaging fluid shear or gas bubbles. Unlike other cell culture systems, such as spinners, hollow-fiber bioreactors, and roller bottles, scale-up is simple, and has been demonstrated up to 100 L of culture volume. The bioreactor is disposable, and therefore requires no cleaning or sterilization. Additions and sampling are possible without the need for a laminar flow cabinet. The unit can be placed in an incubator requiring minimal instrumentation. These features dramatically lower the purchase cost, and operating expenses of this laboratory/pilot scale cell cultivation system. Results are presented for various model systems: 1) recombinant NS0 cells in suspension; 2) adenovirus production using human 293 cells in suspension; 3) Sf9 insect cell/baculovirus system; and 4) human 293 cells on microcarrier. These examples show the general suitability of the system for cells in suspension, anchorage-dependent culture, and virus production in research and GMP applications.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008025016272
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  • 10
    Electronic Resource
    Electronic Resource
    Enhanced CEA production associated with aspirin in a culture of CW-2 cells on some polymeric films (1999)
    Springer
    Cytotechnology 31 (1999), S. 233-242 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; carcinoembryonic antigen ; aspirin ; enhanced production ; Langmuir-Blodgett film
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human colorectal adenocarcinoma tumor (CW2) cells were cultivated in RPMI 1640 media containing 0–7.5 mM aspirin and 10% fetal bovine serum for the production of carcinoembryonic antigen (CEA). By adding aspirin to the media, the production of CEA per cell increased by up to one hundred fold compared to cultivation in normal media containing no aspirin, even though the total cell concentration decreased with the increase in aspirin in the media. The production of CEA was also investigated for CW2 cells cultured on silk fibroin, poly(γ-benzyl-L-glutamate) and poly(γ-benzyl-L-glutamate)/poly(ethylene oxide) diblock copolymer films prepared by the Langmuir-Blodgett and casting methods. The highest production of CEA per cell was observed for the CW2 cells on poly(γ-benzyl-L-glutamate) and its diblock copolymer films prepared by the Langmuir-Blodgett method in the medium containing 5 mM aspirin after 168 hr of inoculation. This originates from the fact that the cell density on the films in the medium containing 5 mM aspirin was the lowest under these conditions. It is suggested that CW2 cells produce CEA more effectively when the cell growth is suppressed by addition of toxic chemicals such as aspirin or by culture on unfavorable films for cell growth.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008030730814
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