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Death rate in a small air-lift loop reactor of vero cells grown on solid microcarriers and in macroporous microcarriers

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Abstract

The death rate of Vero cells grown on Cytodex-3 microcarrierswas studied as a function of the gas flow rate in a smallair-lift loop reactor. The death rate may be described byfirst-order death-rate kinetics. The first-order death-rateconstant as calculated from the decrease in viable cells, theincrease in dead cells and the increase in LDH activity islinear proportional to the gas flow rate, with a specifichypothetical killing volume in which all cells are killed ofabout 2.10-3m3 liquid per m3 of air bubbles.In addition, an experiment was conducted in the sameair-lift reactor with Vero cells grown inside porous Asahimicrocarriers. The specific hypothetical killing volumecalculated from this experiment has a value of 3.10-4m3 liquid per m3 of air bubbles, which shows thatthe porous microcarriers were at least in part able to protectthe cells against the detrimental hydrodynamic forcesgenerated by the bubbles.

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References

  • Bavarian, F, Fran LS, Chalmers J (1991) Microscopic visualization of insect cell bubble interactions. I: Rising bubbles, air-medium interface, and the foam layer. Biotechnol Prog 7, 140–150.

    Google Scholar 

  • Beverloo WA and Tramper J (1994) Intensity of microcarrier collisions in turbulent flow. Bioproc Eng 11, 177–184.

    Google Scholar 

  • Bugarski B, King GA, Daugulis AJ and Goosen MFA (1989) Performance of an external air-lift bioreactor for the production of monoclonal antibodies by immobilized hybridoma cells. Applied Microbiol Biotechnol 30, 264–269.

    Google Scholar 

  • Cahn F (1990) Biomaterials aspects of porous microcarriers for animal cell culture. Tibtech 8, 131–136.

    Google Scholar 

  • Chalmers JJ and Bavarian F (1991) Microscopic visualization of insect cell-bubble interactions. II: The bubble film and bubble rupture. Biotechnol Prog 70, 151–158.

    Google Scholar 

  • Cherry RS and Papoutsakis ET (1986) Hydrodynamic effects on cells in agitated tissue culture reactors. Bioproc Eng 1, 29–41.

    Google Scholar 

  • Cherry RS and Hulle CT (1992) Cell death in the films of bursting bubbles. Biotechnol Prog 8, 11–18.

    Google Scholar 

  • Christi MY (1989) Airlift bioreactors. Elsevier Science Publishers LTD, New York.

    Google Scholar 

  • Croughan MS, Sayre ES and Wang DIC (1989) Viscous reduction of turbulent damage in animal cell culture. Biotechnol Bioeng 33, 862–872.

    Google Scholar 

  • Croughan MS, Wang DIC (1990) Reversible removal and hydrodynamic phenomena in CHO microcarrier cultures. Biotechnol Bioeng 38, 316–319.

    Google Scholar 

  • Gardner AR, Gainer JL and Kirwan DJ (1990) Effects of stirring and sparging on cultured hybrisoma cells. Biotechnol Bioeng 35, 940–947.

    Google Scholar 

  • Geaugey V, Pascal F, Engasser JM and Marc A (1990) Influence of the culture oxygenation on the release of LDH by hydridoma cells. Biotechnol Techn 4, 257–262.

    Google Scholar 

  • Handa-Corrigan A, Emery AN and Spier RE (1989) Effect of gasliquid interfaces on the growth of suspended mammalian cells: mechanisms of cell damage by bubbles. Enzyme Microb Technol 11, 230–235.

    Google Scholar 

  • Jöbses I, Martens D and Tramper J (1991) Lethal events during gas sparging in animal cell culture. Biotechnol Bioeng 37, 484–490.

    Google Scholar 

  • Jordan M, Sucker H, Einsele F and Eppenberger HM (1994) Interactions between animal cells and gas bubbles: The influence of serum and pluronic F68 on the physical properties of the bubble surface. Biotechnol Bioeng 43, 446–454.

    Google Scholar 

  • Kunas KT and Papoutsakis ET (1990) Damage mechanisms of suspended animal cells in agitated bioreactors with and without bubble entrainment. Biotechnol Bioeng 36, 476–483.

    Google Scholar 

  • Lakhotia S and Papoutsakis ET (1992) Agitation induced cell injury in microcarrier cultures. Protective effect of viscosity is agitation intensity dependent: Experiments and modelling. Biotechnol Bioeng 39, 95–107.

    Google Scholar 

  • Legrand C, Bour JM, Capiaumont J, Martial A, Marc A, Wudtke M, Kretzmer G, Demangel C, Duval D and Hache J (1992) Lactate dehydrogenase (LDH) activity of the number of dead cells in the medium of cultured eukaryotic cells as marker. J Biotechnol 25, 231–243.

    Google Scholar 

  • Lu GZ, Thompson BG and Gray MR (1992) Physical modelling of animal cell damage by hydrodynamic forces in suspension cultures. Biotechnol Bioeng 40, 1277–1281.

    Google Scholar 

  • Ludwig A, Kretzmer G and Schügerl K (1992) Determination of a 'critical shear stress level' applied to adherent mammalian cells. Enzyme Microb Technol 14, 209–213.

    Google Scholar 

  • Martens DE, Gooijer CD de, Beuvery EC and Tramper J (1992) Effect of serum concentration on hybridoma viable cell density and production of monoclonal antibodies in CSTRs and on shear sensitivity in air-lift loop reactors. Biotechnol Bioeng 39, 891–897.

    Google Scholar 

  • Martens DE, Gooijer CD de, Velden-de Groot CAM, van der Beuvery EC and Tramper J (1993) Effect of dilution rate on growth, productivity, cell cycle and size, and shear sensitivity of a hybridoma cell in a continuous culture. Biotechnol Bioeng 41, 429–439.

    Google Scholar 

  • Michaels JD, Petersen JF, McIntire LV and Papoutsakis ET (1991) Protection mechanisms of freely suspended animal cells (CRL 8018) from fluid-mechanical injury. Viscometric and biorector studies using serum, pluronic F68 and polyethylene glycol. Biotechnol Bioeng 38, 169–180.

    Google Scholar 

  • Mukhopadhyay A, Mukhopadhyay SN and Talwar GP (1993) Cellular affinity is the only deciding factor for microcarrier selection for the cultivation of anchorage dependent cells. Biotechnol Techn 7, 173–176.

    Google Scholar 

  • Murhammer DW and Goochee CF (1990) Sparged animal cell bioreactors: Mechanism of cell damage and pluronic F-68 protection. Biotechnol Prog 6, 391–397.

    Google Scholar 

  • Nikolai T and Hu W-S (1992) Cultivation of mammalian cells on macroporous microcarriers. Enzyme Microb Technol 14, 203–213.

    Google Scholar 

  • Nollert MU, Diamond SL and McIntire LV (1991) Hydrodynamic shear stress and mass transport modulation of endothelial cell metabolism. Biotechnol Bioeng 38, 588–602.

    Google Scholar 

  • O'Connor KC, Papoutsakis ET (1992) Agitation effects on microcarrier and suspension CHO cells. Biotechnol Techn 6, 323–328.

    Google Scholar 

  • Papoutsakis ET (1991) Fluid mechanical damage of animal cells in bioreactors. TibTech 9, 427–437.

    Google Scholar 

  • Petersen JF, McIntire LV and Papoutsakis ET (1988) Shear sensitivity of cultured hybridoma cells (CRL-81018) depends on mode of growth, culture age and metabolic concentrations. J Biotechnol 7, 229–246.

    Google Scholar 

  • Pol L van der, Zijlstra G, Thalen M and Tramper J (1990) Effect of serum concentration on production of monoclonal antibodies and on shear sensitivity of a hybridoma. Bioproc Eng 5, 241–245.

    Google Scholar 

  • Pol L van der, Bonarius D, Wouw G van de and Tramper J (1993) Effect of silicone antifoam on shear sensitivity of hybridoma cells in sparged cultures. Biotechnol Prog 9, 504–509.

    Google Scholar 

  • Riet K van 't and Tramper J (1991) Basic bioreactor design. Marcel Dekker Inc., New York.

    Google Scholar 

  • Shiragami N, Honda H and Unno H (1993) Anchorage dependent animal cell culture using a porous microcarrier. Bioproc Eng 8, 295–299.

    Google Scholar 

  • Tramper J, Williams JB, Joustra D (1986) Shear sensitivity of insect cells in suspension. Enzyme Microbiol Technol 8, 33–36.

    Google Scholar 

  • Tramper J, Smit D, Straatman J and Vlak JM (1988) Bubble column design for growth of fragile insect cells. Bioproc Eng 3, 37–41.

    Google Scholar 

  • Trinh K, Garcia-Briones M and Chalmers JJ (1994) Quantification of damage to suspended insect cells as a result of bubble rupture. Biotechnol Bioeng 43, 37–45.

    Google Scholar 

Download references

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Author notes

  1. E.A.A. Nollen

    Present address: Department of Radiobiology, University of Groningen, Bloemsingel1, Groningen, The Netherlands

Authors and Affiliations

  1. Department of Food Science, Food and Bioengeneering Group, Agricultural University, Bomenweg 2, 6703 HD, Wageningen, The Netherlands

    D.E. Martens, C.D. de Gooijer & J. Tramper

  2. Laboratory of Inactivated Viral Vaccines, National Institute of Public Health and Environmental Protection, P.O. Box 1, 3720 BA, Bilthoven, The Netherlands

    M. Hardeveld, C.A.M. van der Velden-de Groot & E.C. Beuvery

Authors
  1. D.E. Martens
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  2. E.A.A. Nollen
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  3. M. Hardeveld
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  4. C.A.M. van der Velden-de Groot
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  5. C.D. de Gooijer
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  6. E.C. Beuvery
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  7. J. Tramper
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Martens, D., Nollen, E., Hardeveld, M. et al. Death rate in a small air-lift loop reactor of vero cells grown on solid microcarriers and in macroporous microcarriers. Cytotechnology 23, 61–75 (1997). https://doi.org/10.1023/B:CYTO.0000010399.23373.65

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  • DOI: https://doi.org/10.1023/B:CYTO.0000010399.23373.65

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