, Volume 15, Issue 1–3, pp 311–320 | Cite as

Cells and bubbles in sparged bioreactors

  • Jeffrey J. Chalmers


Ever since animal cells have been grownin-vitro, various techniques have been used to supply the cells with oxygen. The most simple and commonly used ‘large-scale’ technique to provide oxygen is through the introduction of gas bubbles. However, almost since the beginning ofin-vitro cell culture, empirical observations have indicated that bubbles can be detrimental to the cells. This review will discuss the background of the problem, review the relevant research on the topic, attempt to provide a coherent summary of what we know from all of this research, and finally outline what still needs to be investigated. Specific topics to be covered include: experimental correlations of cell damage with bubbles, cell attachment to bubbles, the hydrodynamics of bubble repture, bioreactor studies, visualization studies, and computer simulations and qualification of cell death as a result of bubble rupture.

Key Words

Animal cells bioreactors bubbles damage rupture 


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  1. Augenstein DC, Sinskey AJ, Wang DIC (1971) Effect of shear on the death of two strains of mammalian tissue cells. Biotechnol. Bioeng. 13: 409–418.Google Scholar
  2. Aunins JG, Croughan MS, Wang DIC (1986) Engineering developments in homogenous culture of animal cells: Oxygenation of reactors and scaleup. Biotechnol. Bioeng. 17: 399–723.Google Scholar
  3. Backer MP, Metzger LS, Slaber PL, Nevitt KL, Boder GB (1988) Large-scale production of nonoclonal antibodies in suspension culture. Biotechnol. Bioeng. 32: 993–1000.Google Scholar
  4. Bavarian F, Fan LS, Chalmers JJ (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
  5. Boulton-Stone JM, Blake JR (1993) Gas-bubbles bursting at a free surface. J. Fluid Mech. 154: 437–466.Google Scholar
  6. Cherry RS, Papoutsakis ET (1986) Hydrodynamic effects on cells in agitated tissue culture reactors. Bioproc. Eng. 1: 29–41.Google Scholar
  7. Chalmers JJ, Bavarian F (2991) Microscopic visualization of insect cell-bubble interactions. II: The bubble film and bubble rupture. Biotechnol. Prog. 7: 151–158.Google Scholar
  8. Croughan MS, Hamel JF, Wang DIC (1987) Hydrodynamic effects in animal cells grown in microcarrier cultures. Biotechnol. Bioeng. 29: 130–141.Google Scholar
  9. Dodge TC, Hu WS (1986) Growth of hybridoma cells under different agiatation conditions. Biotechnol. Letters 8: 683–686.Google Scholar
  10. Garcia-Briones MA, Brodkey RS, Chalmers JJ (1994) Computer simulations of the rupture of a gas bubble at a gas-liquid interface and its implications in animal cell damage. Chem. Eng. Sci. 49: 2301–2320.Google Scholar
  11. Garcia-Briones MA, Chalmers JJ (1992) Cell-bubble interactions: Mechanims of suspended cell damage. Ann. N.Y, Acad. Sci. 665: 219–229.Google Scholar
  12. Goldblum S, Bae Y, Hink WF, Chalmers JJ (1990) Protective effect of methylcellulose and other polymers on insect cells subjected to laminar shear stress. Biotechnol. Prog. 6: 383–390.Google Scholar
  13. Handa-Corrigan A, Emary AN, Spier RE (1989) Effect of gas-liquid interfaces on the growth of suspended mammalian cells: mechanisms of cell damage by bubbles. Enzyme Microb. Technol. 11: 230–235.Google Scholar
  14. Handa A, Emary AN, Spier RE (1987) On the evaluation of gasliquid interfacial effects on hydridoma viability in bubble column bioreactors. Dev. Biol. Stand. 66: 241–253.Google Scholar
  15. Hu WS, Meier J, Wang DIC (1985) A mechanistic analysis of the inoculum requirement for the cultivation of mammalian cells on microcarriers. Biotechnol. Bioeng. 27: 585–595.Google Scholar
  16. Jobses I, Martens D, Tramper J (1991) Lethal events during gas sparging in animal cell culture. Biotech. Bioeng. 37: 484–490.Google Scholar
  17. Kilburn DG, Webb FC (1968) The cultivation of animal cells at controlled dissolved oxygen partial pressure. Biotechnol. Bioeng. 10: 801–814.Google Scholar
  18. Kunas KT, Papoutsakis ET (1990a) Damage mechanisms of suspended animal cells in agitated bioreactors with and without bubble entrainment. Biotechnol. Bioeng. 36: 476–483.Google Scholar
  19. Kunas KT, Papoutsakis ET (1990b) The protective effect of serum against hydrodynamic damage of hydridoma cells in agitated and surface-areated bioreactors. J. Biotechnol. 15: 57–70.Google Scholar
  20. Lee GM, Huard TK, Kaminski MS, Palsson BO (1988) Effect of mechanical agitation on hydridoma cell growth. Biotechnol. Letters 10: 625–628.Google Scholar
  21. MacIntyre F (1972) Flow patterns in breaking bubbles. J Geophys. Res. 77: 5211–5228.Google Scholar
  22. MacIntyre F (1968) Bubbles: a boundary-layer ‘microtom” for micron-thick samples of a liquid surface. J. Phys. Chem. 72: 589–592.Google Scholar
  23. Martens DE, de Gooijer CD, Beuvery EC, 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
  24. Murhammer DW, Goochee CF (1990) Sparged animal cell bioreactors: mechanism of cell damage and Pluronic F-68 protection. Biotechnol. Prog. 6: 391–397.Google Scholar
  25. Oh SKW, Nienow AW, Al-Rubeai M, Emary AN (1989) The effect of agiatation intensity with and without continuous sparging on the growth and antibody production of hybridoma cells. J. Biotechnol. 12: 45–62.Google Scholar
  26. Orton D, Wang DIC (1991) Fluorescent Visualization of Cell Death in Bubble Areated Bioreactors. Cell Culture Engineering III, Engineering Foundation, Feb. 2–7,Google Scholar
  27. Ruyan WS, Gyer RP (1963) Growth of L cell suspensions on a Warburg apparatus. Proc. Soc. Bio. Med. 103: 252–254.Google Scholar
  28. Schurch U, Kramer H, Einsle A, Widmer F, Eppenberger HM (1988) Experimental evaluation of laminar shear stress on the behaviour of hybridoma mass cell cultures producing monoclonal antibodies against mitochondrial creatine kinase. J. Biotechnol. 7: 179–184.Google Scholar
  29. Sinskey AJ, Fleischaker RJ, Tyo MA, Giard DJ, Wang DIC (1981) Production of cell derived products: virus and interferon. Ann. N.Y. Acad. Sci. 369: 47–59.Google Scholar
  30. Smith CG, Greenfield PF, Randerson DH (1987) A technique for determining the shear sensitivity of mammalian cells in suspension culture. Biotechnol. Techn. 1: 39–44.Google Scholar
  31. Swim HE, Parker RF (1960) Effect of Pluronic F-68 on growth of fibroblasts in suspension on rotary shakers. Proc. Soc. Biol. Med. 103: 252–254.Google Scholar
  32. Tramper J, Smit JD, Straatman J, Valk JM (1988) Bubble-column design for growth of fragile insect cells. Bioprocess Engin. 3: 37–41.Google Scholar
  33. Tramper J, Williams JB, Joustra D (1986) Shear sensitivity of insect cells in suspension. Enzyme Microb. Technol. 8: 33–36.Google Scholar
  34. Trinh K, Garcia-Briones MA, Hink FH, Chalmers JJ (1994) Quantification of damage to suspended insect cells as a result of bubble rupture. Biotechnol. Bioeng. 43: 37–45.Google Scholar
  35. Wang NS, Yang JD, Calabrese RV, Chang KC (1994) Unified modeling framework of cell death due to bubbles in agitated and sparged bioreactors. J. Biotechnol. 33: 107–122.Google Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Jeffrey J. Chalmers
    • 1
  1. 1.Department of Chemical EngineeringOhio State UniversityColumbusUSA

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