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Shear sensitivity of insect cells

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Conclusions

While insect cells can be easily damaged in bioreactors as a result of hydrodynamic forces, it is also relatively easy to prevent this damage. Of several possible damage mechanisms, the best understood and preventable is the attachment of cells to gas-liquid interfaces and the subjection of these attached cells to the hydro-dynamic forces and/or physical forces associated with these interfaces. For example, cells attached to gas bubbles in a bioreactor can be transported into the foam layer where they are physically removed from the cell suspension, or they can be killed when the gas bubble they are attached to ruptures at the medium-air interface at the top of the bioreactor. The easiest method to prevent this damage is through the use of specific surface active compounds, such as Pluronic F-68 or Methocel E-50 which prevent the cells from attaching to the gas-medium interface.

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References

  • AugensteinDC, SinskeyAJ & WangDIC (1971) Effect of shear on the death of two strains of mammalian tissue cells. Biotechnol. Bioeng. 13: 409–418.

    Google Scholar 

  • AgathosSN, JeongY-H & VenkatK (1990) Growth kinetics of free and immobilized insect cell cultures. Ann. N.Y. Acad. Sci. 589: 372–398.

    Google Scholar 

  • Binh J, Jarnagin K, Williams S, Chan H & Barnett J (0000) Fed-batch culture of insect cells: a method to increase the yield of recombinant human nerve growth factor (rhNGF) in the baculovirus expression system. J. Biotechnol. 31: 205–217.

    Google Scholar 

  • BackerM, MetzgerL, SlaberP, NevittK & BoderG (1988) Largescale production of monoclonal antibodies in suspension culture. Biotechnol. Bioeng. 32: 993–1000.

    Google Scholar 

  • BavarianF, FanLS & ChalmersJJ (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 

  • Boulton-StoneJM & BlakeJR (1993) Gas-bubbles bursting at a free surface. J. Fluid Mech. 154: 437–466.

    Google Scholar 

  • CameronIR, PosseeRD, BishopDHL, (1989) Insect cell culture technology in baculovirus expression systems. TIBTECH 7: 66–70.

    Google Scholar 

  • CaronAW, ArchambaultJ & MassieB (1990) High level recombinant protein production in bioreactors using the baculovirus-insect cell expression system. Biotechnol. Bioeng. 36: 1133–1140.

    Google Scholar 

  • CaronAW, TomRL, KamenAA & MassisB (1994) Baculovirus expression systems scaleup by perfusion of high-density SF-9 cell cultures Biotechnol. Bioeng. 43: 881–891.

    Google Scholar 

  • ChalmersJJ & BavarianF (1991) Microscopic visualization of insect cell-bubble interactions. II: The bubble film and bubble rupture. Biotechnol. Prog. 7: 151–158.

    Google Scholar 

  • ChattopadhyayD, RathmanJF & ChalmersJJ (1995a) The protective effect of specific medium additves with respect to bubble rupture. Biotechnol. Bioeng. 45: 473–480.

    Google Scholar 

  • ChattopadhyayD, RathmanJF & ChalmersJJ (1995b) Thermodynamic Approach to Explain Cell Adhesion to Air-Medium Interfaces. Biotechnol: Bioeng. 48: 649–658.

    Google Scholar 

  • DeutschmannSM & JaegerV (1994) Optimization of the growth conditions of Sf21 insect cells for high-density perfusion culture in stirred-tank bioreactor. Enzyme Microb. Technol. 16: 506–512.

    Google Scholar 

  • DodgeT & HuW (1986) Growth of hybridoma cells under different agitation conditions, Biotechnol. Letters. 8: 683–686.

    Google Scholar 

  • FowlerMW (1984) Plant cell culture: natural products and industrial applications. Genet. Eng. Rev. 2: 41–67.

    Google Scholar 

  • Garcia-BrionesMA, BrodkeyRS & ChalmersJJ (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 

  • Garcia-BrionesMA & ChalmersJJ (1992) Cell-bubble interactions: Mechanisms of suspended cell damage, Ann. N.Y. Acad. Sci. 665: 219–229.

    Google Scholar 

  • GlackenMW, FlesichakerRJ & SinskeyAJ (1983) Mammalian cell culture: engineering principles and scale-up, Trends Biotechnol. 1: 102–108.

    Google Scholar 

  • GoldblumS, BaeY, HinkWF & ChalmersJJ (1990) Protective effect of methylcellulose and other polymers on insect cells subjected to laminar shear stress. Biotechnol. Prog. 6: 383–390.

    Google Scholar 

  • Handa-CorriganA, EmaryAN & SpierRE (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 

  • HandaA, EmeryA & SpierRE (1987) On the evaluation of gas-liquid interfacial effects on hybridoma viability in bubble column bioreactors, Develop. Biol. Standard 66: 241–252.

    Google Scholar 

  • HinkFH (1982) Production of Autographa Californica Nuclear Polyhedrosis virus in cell from large-scale suspension cultures, In: Microbial and Viral Pesticides, KurstakE (ed.) Marcel Dekker, NY.

    Google Scholar 

  • HinkWF & StraussE (1979), Suspension culture of the Cabbage Looper (TN-368) cell line, Tissue Cult. Assoc. Man. 5: 1023–1025.

    Google Scholar 

  • HulscherM & OnkenU (1988), Influence of bovine serum albumin on the growth of hybridoma cells in air-lift loop reactors using serum free medium, Biotechnol. Lett. 10: 689–694.

    Google Scholar 

  • JobsesI, MartensD & TramperJ (1990) Lethal effects during gas sparging in animal cell culture. Biotechnol. Bioeng. 10: 801–814.

    Google Scholar 

  • KamenAA, TomRL, CaronAW, ChavarieC, MassieB & ArchambaultJ (1991) Culture of insect cells in a helical ribbon impeller bioreactor, Biotechnol. Bioeng. 38: 619–628.

    Google Scholar 

  • KilburnDG & WebbF (1968) The cultivation of animal cells at controlled dissolved oxygen partial pressure, Biotechnol. Bioeng. 10: 801–814.

    Google Scholar 

  • KimHR, LeeKW, KimTY, OhJH, YangJM, KangSK & ChungIS (1994) Insect Cell Culture for recombinant β-galactosidase production using a spin-filter bioreactor. J. Microbiol. Biotechnol. 4: 200–203.

    Google Scholar 

  • KunasKT & PapoutsakisET (1990a) The protective effect of serum against hydrodynamic damage of hybridoma cells in agitated and surface-aerated bioreactors, J. Biotechnol. 15: 57–70.

    Google Scholar 

  • KunasKT & PapoutsakisET (1990b) Damage mechanism of suspended animal cells in agitated bioreactors with and without bubble entrainment, Biotechnol. Bioeng. 36: 476–483.

    Google Scholar 

  • LeeG, HuardT, KaminskiM & PalssonB (1988) Effect of mechanical agitation on hydridoma cell growth. Biotechnol. Letters 10: 625–628.

    Google Scholar 

  • LengyelJ, SpradlingA & PenmanS (1975) Methods with insect cells in suspension culture II Drosophila melanogaster. In: Methods of Cell Biology, Vol. 10, PrescottDM (ed.) Academic Press, NY.

    Google Scholar 

  • MacIntyreF (1968) Bubbles: a boundary-layer “microtome” for micron thick samples of a liquid surface, J. Phys. Chem. 72: 589–592.

    Google Scholar 

  • MacIntyreF (1972) Flow patters in breaking bubbles, J. Geophys. Res. 77: 5211–5228.

    Google Scholar 

  • MartensDE, deGooijerCD, BeuveryEC & TramperJ (1992) Effect of serum on hybridoma viable cell density and production of monoclonal antibodies in CSTR's and on shear sensitivity in airlift loop reactors. Biotechnol. Bioeng. 39: 891–897.

    Google Scholar 

  • MaiorellaB, InlowD, ShaugerA & HaranoD (1988) Large scale insect cell-culture for recombinant protein production. Bio/technol. 6, 1406–1410.

    Google Scholar 

  • MichaelsJD, & PapoutsakisEF (1991) Polyvinyl alcohol and polyethylene glycol as protectants against fluid-mechanical injury of freely-suspended animal cells. J. Biotechnol. 19: 241–258.

    Google Scholar 

  • MichaelsJD, KunasKT & PapoutsakisET (1992) Fluid Mechanical damage of freely suspended animal cells in agitated bioreactors: Effects of dextran, dreivatized celluloses, and polyvinyl alcohol. Chem. Eng. Comm. 118: 341–360.

    Google Scholar 

  • MichaelsJD, NowakJE, MallikAK, KoczoK, WasanDT & PapoutsakisET (1995a) Analysis of Cell-to-Bubble Attachment in Sparged Bioreactors in the Presence of Cell-Protecting Additives. Biotechnol. Bioeng. 47: 407–419.

    Google Scholar 

  • MichaelsJD, NowakJE, MallikAK, KoczoK, WasanDT & PapoutsakisET (1995b) Interfacial Properties of Cell Culture Media with Cell-Protecting Additives. Biotechnol. Bioeng. 47: 420–430.

    Google Scholar 

  • MiyakeT, SaigoK, MarunouchiT & ShibaT (1977) Suspension culture of Drosophila cells employing a gyratory shaker, In Vitro, 13: 245–251.

    Google Scholar 

  • MizrahiA (1984), Oxygen in human lymphoblastoid cell line cultures and effect of polymers in agitated and aerated cultures, Devel Biol Std 55: 93–102.

    Google Scholar 

  • MurhammerDW & GoocheeCF (1988) Scaleup of insect cell cutlures: protective effects of Pluronic F-68. Bio/technol. 6: 1411–1415.

    Google Scholar 

  • MurhammerDW & GoocheeCF (1990) Sparged animal cell bioreactors: mechanisms of cell damage and Pluronic F-68 protection. Biotechnol. Prog. 6: 391–397.

    Google Scholar 

  • OhSKW, NienowAW, Al-Rubeai & EmeryAN (1989) The effect of agitation intensity with and without continuous sparging on the growth and antibody production of hybridoma cells. J. Biotechnol. 12: 45–62.

    Google Scholar 

  • OhSKW, NienowAW, Al-RubeaiM & EmeryAN (1992) Further studies on the culture of mouse hybridomas in an agitated bioreactor with and without continuous sparging. J. Biotechnol. 22: 245–270.

    Google Scholar 

  • Orton D & Wang DIC (1991) Fluorescent visualization of cell death in bubble areated bioreactors, Cell Culture Engineering III, Engineering Foundation, Feb. 2–7.

  • RunyanW & GyerR (1963) Growth of L cell suspensions on a Warburg apparatus Proc. Soc. Biol. Med. 112: 1027–1030.

    Google Scholar 

  • SchurchU, KramerH, EinsleA, WidmerF & EppenbergerHM (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 

  • SilvaHJ, CortinasT & ErtolaRJ (1987) Effect of hydrodynamic stress on Dunaliella growth. J. Chem. Technol. Biotechnol. 40: 41–49.

    Google Scholar 

  • SmithCG, GreenfieldPF & RandersonDH (1987) A technique for determining the shear sensitivity of mammalian cells in suspension culture. Biotechnol. Tech. 1: 39–44.

    Google Scholar 

  • SpradlingA, StingerRH, LengyelJ & PenmanS (1975) Methods with insect cells in suspension culture. I. Aedes albopictus. In: Methods of Cell Biology, Vol. 10, PrescottDM (ed.) Academic Press, NY.

    Google Scholar 

  • SwimH & ParkerR (1960) Effect of Pluronic F-68 on growth of fibroblasts in suspension on rotary shakers. Proc. Soc. Biol. Med. 103: 252–254.

    Google Scholar 

  • TellingRC & ElsworthR (1965) Submerged culture of hamster kidney cells in a stainless steel vessel, Biotechnol. Bioeng. 7: 417–434.

    Google Scholar 

  • TramperJ, WilliamsJ & JoustraD, (1986), Shear sensitivity of insect cells in suspension, Enzyme Microb. Technol. 8: 33–36.

    Google Scholar 

  • TrinhK, Garcia-BrionesMA, HinkFH & ChalmersJJ (1994), Quantification of damage to suspended insect cells as a result of bubble rupture, Biotechnol. Bioeng. 43: 37–45.

    Google Scholar 

  • Vaughn JL (1967) Growth of insect cell lines in suspension culture, In: Proc. Int. Colloq Invertebr. Tissue Cult. 2nd Tremezzo Como, Italy. Wang NS, Yang JD, Calabrese RV, Chang KC (1994) Univfied modeling framework of cell death due to bubbles in agitated and sparged biorectors. J. Biotechnol. 33: 107–122.

    Google Scholar 

  • YangJD & WangNS (1992) Cell Inactivation in the Presence of Sparging and Mechanical Agitation, Biotechnol. Bioeng. 40: 806–816.

    Google Scholar 

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Chalmers, J.J. Shear sensitivity of insect cells. Cytotechnology 20, 163–171 (1996). https://doi.org/10.1007/BF00350397

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