FLUID-MECHANICAL DAMAGE OF FREELY-SUSPENDED ANIMAL CELLS IN AGITATED BIOREACTORS: EFFECTS OF DEXTRAN, DERIVATIZED CELLULOSES AND POLYVINYL ALCOHOL

Abstract Different media additives were used to analyze cell damage and “shear protection” characteristics of freely suspended animal cells (CRL-8018) cultured in stirred tank bioreactors. The additives used included dextran, polyvinyl alcohol (PVA), and methylcellulose, which are viscosity enhancers, surface active agents or combinations of these, respectively. We used these additives to study the effects of bulk viscosity and interfacial properties on cell damage or protection under conditions whereby bubble-induced cell damage becomes inhibitory to cell growth if the medium is not supplemented with a protective additive. Dextran (229 kDa, 1–3% w/v) was used to analyze the effects of increased bulk viscosity on cell damage or protection. It was found that the presence of dextran increased cell death under intense agitation. Several grades and types of derivatized methylcelluloses including Methocel A15LV (15 kDa, 0.1–0.5 w/v%), Methocel E50LV (50 kDa, 0.1% w/v), and Methocel H100LV (100 kDa, 0.25% w/v),...

[1]  J. Bryant MAMMALIAN CELLS IN CHEMICALLY DEFINED MEDIA IN SUSPENSION CULTURES * , 1966, Annals of the New York Academy of Sciences.

[2]  W. F. Hink,et al.  Protective Effect of Methylcellulose and Other Polymers on Insect Cells Subjected to Laminar Shear Stress , 1990, Biotechnology progress.

[3]  J. Tramper,et al.  Some Engineering and Economic Aspects of Continuous Cultivation of Insect Cells for the Production of Baculoviruses , 1986 .

[4]  E. Papoutsakis,et al.  Polyvinyl alcohol and polyethylene glycol as protectants against fluid-mechanical injury of freely-suspended animal cells (CRL 8018). , 1991, Journal of biotechnology.

[5]  J. M. Vlak,et al.  Bubble-column design for growth of fragile insect cells , 1988 .

[6]  W. Runyan,et al.  Growth of L Cell Suspensions on a Warburg Apparatus.∗ , 1963, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[7]  F. Bavarian,et al.  Microscopic Visualization of Insect Cell‐Bubble Interactions. II: The Bubble Film and Bubble Rupture , 1991, Biotechnology progress.

[8]  Raymond E. Spier,et al.  Effect of gas—liquid interfaces on the growth of suspended mammalian cells: mechanisms of cell damage by bubbles , 1989 .

[9]  D. J. Merchant,et al.  The mechanism of cell binding and cell-sheet formation in L strain fibroblasts. , 1960, Experimental cell research.

[10]  J. Tramper,et al.  Shear sensitivity of insect cells in suspension , 1986 .

[11]  F. Macintyre,et al.  Flow patterns in breaking bubbles , 1972 .

[12]  Fred E. C. Culick,et al.  Comments on a Ruptured Soap Film , 1960 .

[13]  E. Papoutsakis,et al.  Protection mechanisms of freely suspended animal cells (CRL 8018) from fluid‐mechanical injury. Viscometric and bioreactor studies using serum, pluronic F68 and polyethylene glycol , 1991, Biotechnology and bioengineering.

[14]  R. F. Parker,et al.  Effect of Pluronic F68 on Growth of Fibroblasts in Suspension on Rotary Shaker , 1960, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[15]  R. Spier,et al.  On the evaluation of gas-liquid interfacial effects on hybridoma viability in bubble column bioreactors. , 1987, Developments in biological standardization.

[16]  E. Papoutsakis,et al.  Agitation induced cell injury in microcarrier cultures. Protective effect of viscosity is agitation intensity dependent: Experiments and modeling , 1992, Biotechnology and bioengineering.

[17]  E. Papoutsakis,et al.  Fluid-mechanical damage of animal cells in bioreactors. , 1991, Trends in biotechnology.

[18]  L. Fan,et al.  Microscopic Visualization of Insect Cell‐Bubble Interactions. I: Rising Bubbles, Air‐Medium Interface, and the Foam Layer , 1991, Biotechnology progress.

[19]  Aniruddha B. Pandit,et al.  Mechanically agitated gas-liquid reactors , 1982 .

[20]  J Tramper,et al.  Lethal events during gas sparging in animal cell culture , 1991, Biotechnology and bioengineering.

[21]  G. Moore,et al.  Partial substitution of serum in hematopoietic cell line media by synthetic polymers. , 1970, Applied microbiology.

[22]  E. Papoutsakis,et al.  Media additives for protecting freely suspended animal cells against agitation and aeration damage. , 1991, Trends in biotechnology.

[23]  F. Franek,et al.  Serum-free medium for hybridoma and parental myeloma cell cultivation. , 1986, Methods in enzymology.

[24]  E. Papoutsakis,et al.  Physical mechanisms of cell damage in microcarrier cell culture bioreactors , 1988, Biotechnology and bioengineering.

[25]  E. Papoutsakis,et al.  Damage mechanisms of suspended animal cells in agitated bioreactors with and without bubble entrainment , 1990, Biotechnology and bioengineering.

[26]  T. W. F. Russell,et al.  The design of gas sparged devices for viscous liquid systems , 1978 .

[27]  R. Telling,et al.  Improvements in the growth of BHK-21 cells in submerged culture. , 1971, Applied microbiology.

[28]  Daniel I. C. Wang,et al.  Viscous reduction of turbulent damage in animal cell culture , 1989, Biotechnology and bioengineering.

[29]  Larry V. McIntire,et al.  Shear sensitivity of cultured hybridoma cells (CRL-8018) depends on mode of growth, culture age and metabolite concentration , 1988 .

[30]  R. Elsworth,et al.  Submerged culture of hamster kidney cells in a stainless steel vessel , 1965 .