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One consequence of using low aeration rates to prevent bubble-associated cell damage is the possible accumulation of dissolved CO2 with increasing hydrostatic pressure (or scale) to inhibitory or metabolism-altering concentrations, due to insuf cient ventilation as a result of low air sparging rates or using oxygenenriched air This can be a major problem in large scale bioreactors, especially when operated at high cell densities CO2 production rates between 017 to 13 10 8 mg/cell per hour have been reported for a range of cell lines (Aunins and Henzler, 1993) Kimura and Miller (1996) reported that CHO cells cultivated in laboratory scale bioreactors were moderately tolerant to dissolved CO2 (up to 18% CO2 ), while Gray et al (1996) reported an inhibitory effect at 14% dissolved CO2 in large scale cultures.

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The basic structure for all HTML documents is the same and should include the following minimum elements and tags: <DOCTYPE> The declared type of the document <html> The main container for HTML pages <head> The container for page header information <title> The title of the page <body> The main body of the page These elements t together in the following template format:

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CO2 ventilation rate is affected by control strategy used to control dissolved oxygen (surface aeration, air sparging, partial pressure of oxygen in the inlet gas, and type of sparger) and therefore, a balance between oxygenation, CO2 ventilation, and pH control needs to be made One solution to this problem is to employ two spargers serving different functions; a sintered sparger to provide small bubbles to enhance oxygen mass transfer and a pipe sparger producing relatively larger bubbles for CO2 ventilation while ensuring minimal cell damage The inhibitory effects of CO2 are evident at lower concentrations if the culture osmolality is >300 mOs, as may be the case in fed-batch cultures Increased osmolality up to 400 mOs has also been shown to increase the speci c antibody production rate, although this enhancement is cellline dependent.

De nitions In Europe in 1999, probiotics accounted for more than two-thirds of the total market for functional foods. They are live cultures of micro-organisms, usually bacteria that survive passage through the upper parts of the gut, particularly the acid environment of the stomach, and adhere to and colonise the bowel where they favourably alter the microbial balance. When they colonise the bowel they displace other potentially pathogenic bacteria

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Cyclic exposure of cells to increasing hydrostatic pressure with scale can also occur, although results reported by Tagaki et al (1995) at constant pH, pCO2 , and pO2 using hybridoma cultures suggested that this is unlikely to affect culture metabolism per se Despite advances in our understanding of cell damage due to bubble aeration and under realistic conditions of agitation, we still lack suf cient knowledge for an optimal a priori design This is due to incomplete understanding of the complex two-phase hydrodynamics in bioreactors (eg, bubble size distribution in large scale cell culture media), bubble cell attachment mechanisms, effects of surfactants on media physical properties and due to the diversity of cell line, type of culture media, and operating conditions.

However, a general guideline for design and operation of large scale bioreactors for freely suspended animal cells should probably consider the following (Nienow, 1997): Multiple impellers in high aspect ratio bioreactors with a large diameter radial ow Rushton turbine or Scaba (or up-pumping axial ow) impeller with clearance ratios between 033 and 05, good for gas dispersion, and an up-pumping or down-pumping large diameter axial ow impeller for liquid blending above it Increased agitation intensity to ensure good liquid blending is also recommended Design and operating conditions should be chosen to avoid pH excursions Clean in place issues related to addition pipes for subsurface addition of concentrated reagents should be addressed..

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