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  • 1
    Electronic Resource
    Electronic Resource
    The effect of Pluronic F-68 on hybridoma cells in continuous culture (1992)
    Springer
    Applied microbiology and biotechnology 37 (1992), S. 44-45 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The comparative effects of Pluronic on cell growth of hybridomas were studied. The addition of 0.05% Pluronic decreased the steady-state cell number in continuous culture by about 12% compared to a non-supplemented culture. In short-term experiments, results demonstrated a gradual decrease in cell number with increasing concentration of Pluronic. Such growth inhibition was found to be a result of lowering the rate of DNA synthesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00174201
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  • 2
    Electronic Resource
    Electronic Resource
    Oxygenation of intensive cell-culture system (1995)
    Springer
    Applied microbiology and biotechnology 43 (1995), S. 1028-1033 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The abilities of various methods of oxygenation to meet the demands of high-cell-density culture were investigated using a spin filter perfusion system in a bench-top bioreactor. Oxygen demand at high cell density could not be met by sparging with air inside a spin filter (oxygen transfer values in this condition were comparable with those for surface aeration). Sparging with air outside a spin filter gave adequate oxygen transfer for the support of cell concentrations above 107 ml−1 in fully aerobic conditions but the addition of antifoam to control foaming caused blockage of the spinfilter mesh. Bubble-free aeration through immersed silicone tubing with pure oxygen gave similar oxygen transfer rates to that of sparging with air but without the problems of bubble damage and fouling of the spin filter. A supra-optimal level of dissolved oxygen (478% air saturation) inhibited cell growth. However, cells could recover from this stress and reach high density after reduction of the dissolved oxygen level to 50% air saturation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00166920
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  • 3
    Electronic Resource
    Electronic Resource
    A rapid method for evaluation of cell number and viability by flow cytometry (1997)
    Springer
    Cytotechnology 24 (1997), S. 161-168 
    Details
    ISSN: 1573-0778
    Keywords: cell number ; flow cytometry ; proliferationassay ; viability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A simple, rapid and reliable method has been developed for assessing the number and viability of cells, as well as cell size, in suspension culture by the use of flow cytometry. Propidium iodide exclusion is used for viability determination and fluorescent beads serve as an internal standard for cell enumeration. The main advantages of this method are its ability to handle a large number of samples with a high degree of precision and its specificity in detecting viable cells quantitatively in a heterogeneous culture of living and dead cells and debris. The method shows only a fraction of the variation found in the haemacytometer/trypan blue counting method due to its very low operator dependence. CHO - Chinese hamster ovary; FCS - Foetal calf serum; FS - Forward scatter light; MTT - 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide; NCS - newborn calf serum; PBS - Phosphate buffered saline; PI - Propidium iodide; SS - Side scatter light.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007910920355
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  • 4
    Electronic Resource
    Electronic Resource
    Electron microscopy of hybridoma cells with special regard to monoclonal antibody production (1990)
    Springer
    Cytotechnology 4 (1990), S. 13-28 
    Details
    ISSN: 1573-0778
    Keywords: monoclonal antibody ; hybridoma ; electron microscopy ; endoplasmic reticulum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Electron microscopy of mouse hybridoma cell lines shows that the major difference between non, low and high producer cell lines is the amount of endoplasmic reticulum. Vesicular-tubular or cavernous structures of endoplasmic reticulum, which can survive long after cell death, are particularly abundant in producer cell lines. Immunogold labelling with anti-mouse IgG reveals that antibodies are predominantly located in these structures. The cell membrane undergoes structural changes during the late stages of batch culture with the disappearance of microvilli and the appearance of blebs and deep indentations. Necrosis disrupts the cytoplasmic structures and the nucleus is last to degrade.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00148807
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  • 5
    Electronic Resource
    Electronic Resource
    Growth and interferon-γ production in batch culture of CHO cells (1994)
    Springer
    Cytotechnology 15 (1994), S. 65-71 
    Details
    ISSN: 1573-0778
    Keywords: Animal cell culture ; interferon ; CHO ; growth rate ; batch culture ; cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The relationship between growth and interferon-γ (IFN-γ) production in the recombinant cell line CHO 320 was studied by varying the foetal calf serum (FCS) concentration. The specific growth rate varied with the initial FCS concentration in a manner which could be well fitted by the Monod model. TheK s and μ max values were found to be 0.771% (v/v) serum and 0.031 h−1 respectively. The average specific IFN-γ production rates during the exponential phase increased with increasing FCS concentration. A good correlation between specific production rate and specific growth rate was found in all phases of the culture except the lag phase and it was clearly demonstrated that IFN-γ production was growth associated. Specific glucose and glutamine utilisation rates were inversely related to specific growth rates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00762380
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  • 6
    Electronic Resource
    Electronic Resource
    Peptone, a low-cost growth-promoting nutrient for intensive animal cell culture (1994)
    Springer
    Cytotechnology 16 (1994), S. 17-26 
    Details
    ISSN: 1573-0778
    Keywords: Cell culture ; peptone ; media ; intensive culture ; hybridoma ; spin-filter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The effect of addition of peptone to serum-free and serum supplemented media for the growth of hybridoma cells in various systems was studied. Supplementation of defined medium with either proteose peptone or meat peptone resulted in significant increases in cell number and specific monoclonal antibody production in batch culture system. Other peptones were either inactive or less effective. In continuous culture, using medium supplemented with new born calf serum, the addition of peptone resulted in 125% and 150% increases in cell and antibody concentrations respectively. Similar increase in cell number (128%) was also obtained in spin-filter perfusion culture when medium was supplemented with peptone. By comparison, the substitution of a defined 1xMEM amino acids mixture resulted in only a 50% increase. At higher perfusion rates the cell number maintained in steady state using peptone supplement could be increased to 1.3×107 cells ml−1 while the serum concentration was reduced from 5% to 1% at a perfusion rate of 2.5 volumes per day.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00761775
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  • 7
    Electronic Resource
    Electronic Resource
    Modified cellular metabolism in hybridomas subjected to hydrodynamic and other stresses (1990)
    Springer
    Biotechnology letters 12 (1990), S. 323-328 
    Details
    ISSN: 1573-6776
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The responses of hybridoma growth and metabolism to hydrodynamic forces and changes in culture temperature have been examined. DNA synthesis is inhibited under conditions of intense hydrodynamic stress; however, cellular metabolic activity is increased perhaps to feed repair mechanisms. Sub-optimal temperature and nutrient deprivation increase cellular susceptibility to hydrodynamic stress, possibly through the inhibition of repair mechanisms. The reduced cell number in conditions of high agitation may be due at least in part to growth inhibition.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01024425
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  • 8
    Electronic Resource
    Electronic Resource
    A study of uninfected and baculovirus-infected Spodoptera frugiperda cells in T- and spinner flasks (1995)
    Springer
    Biotechnology letters 17 (1995), S. 7-12 
    Details
    ISSN: 1573-6776
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Insect cells have been propagated in monolayers in T-flasks or in suspension culture in spinner flasks, the latter being conducted over a range of spinner speeds. In both configurations, the cells were also infected with either wild or recombinant β-galactosidase baculovirus at MOI of 0.1, 1 and 10. The strength of both uninfected and infected cells was also measured by a micro-manipulation technique. No significant difference in growth rate was obtained between monolayer culture and suspension culture at the spinner rate which was optimum for growth. This optimum was quite sharp. At the lowest speeds cells settled, whilst above the optimum speed the spinner action led to significant cell damage. The maximum infectivity was obtained at this optimum speed which also gave maximum survival after infection. There were significant changes of cell survival and infection, even over relatively small changes of speed, and presumably energy dissipation rate. As changes in growth in turbine-agitated bioreactors have been shown to be much less, even when the energy inputs varied by two orders of magnitude, these findings throw doubt on the usefulness of spinner flasks for assessing “shear” sensitivity of cell lines. The percentage of infected cells and β-galactosidase production were significantly lower in the monolayer culture compared to that in the suspension culture at MOI values below 10 pfu/cell. This difference is explained as being due to the reduced movement of released virus particles from infected to non-infected cells in the T-flasks.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00134187
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  • 9
    Electronic Resource
    Electronic Resource
    Use of a spin-filter can reduce disruption of hybridoma cells in a bioreactor (1993)
    Springer
    Biotechnology techniques 7 (1993), S. 351-356 
    Details
    ISSN: 1573-6784
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The possibility of damage to suspended cells arising from the introduction of a spin-filter rotating at high speed is examined experimentally and theoretically. Far from increasing damage, it is shown that a spin-filter, even when rotating at high speed, can markedly improve the survival of cells in a stirred unbuffled bioreactor. Analysis of the shear stress generated at the filter surface shows values that are orders of magnitude below those shown elsewhere to be damaging to cells. The improvement in survival seen in addition of a spin-filter to an impeller-agitated bioreactor are explained in terms of the events associated with the suppression of vortices, and bubble entrainment and disengagement.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00152540
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  • 10
    Electronic Resource
    Electronic Resource
    Production of α-amylase by Bacillus amyloliquefaciens in batch and continuous culture using a defined synthetic medium (1996)
    Springer
    Biotechnology letters 18 (1996), S. 795-800 
    Details
    ISSN: 1573-6776
    Keywords: Bacillus amyloliquefaciens ; α-amylase ; Continuous fermentation ; Defined medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Using a totally defined synthetic medium the effect of lactose and nitrogen on cell physiology and α-amylase production by Bacillus amyloliquefaciens B155 were investigated. Results showed cell growth and α-amylase production patterns to be similar regardless of the limiting nutrient and suggested stationary phase gene control of α-amylase production as opposed to a direct response to nutrient limitation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00127891
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