Effect of Pluronic F68 on Growth of Fibroblasts in Suspension on Rotary Shaker

Swim, H. E.; Parker, Robert G.

doi:10.3181/00379727-103-25477

Cited by 45 publications

(12 citation statements)

References 1 publication

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“…PF-68, an amphiphilic triblock copolymer consisting of poly(propylene oxide) center and two poly(ethylene oxide) tails, has been the primary additive used industrially to protect cells from damage. It should be added that the original selection of PF-68 was an empirical process (Swim and Parker 1960). A number of mechanisms for the protective effects of PF-68 have been proposed which can be categorized into two groups: biological and physical.…”

Section: Cell Damage From Spargingmentioning

confidence: 99%

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

Hu¹,

2011

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Suspension animal cell culture is now routinely scaled up to bioreactors on the order of 10,000 L, and greater, to meet commercial demand. However, the concern of the 'shear sensitivity' of animal cells still remains, not only within the bioreactor, but also in the downstream processing. As the productivities continue to increase, titer of ~10 g/L are now reported with cell densities greater than 2 × 10(7) cells/mL. Such high, and potentially higher cell densities will inevitably translate to increased demand in mass transfer and mixing. In addition, achieving productivity gains in both the upstream stage and downstream processes can subject the cells to aggressive environments such as those involving hydrodynamic stresses. The perception of 'shear sensitivity' has historically put an arbitrary upper limit on agitation and aeration in bioreactor operation; however, as cell densities and productivities continue to increase, mass transfer requirements can exceed those imposed by these arbitrary low limits. Therefore, a better understanding of how animal cells, used to produce therapeutic products, respond to hydrodynamic forces in both qualitative and quantitative ways will allow an experimentally based, higher, "upper limit" to be created to guide the design and operation of future commercial, large scale bioreactors. With respect to downstream hydrodynamic conditions, situations have already been achieved in which practical limits with respect to hydrodynamic forces have been experienced. This review mainly focuses on publications from both the academy and industry regarding the effect of hydrodynamic forces on industrially relevant animal cells, and not on the actual scale-up of bioreactors. A summary of implications and remaining challenges will also be presented.

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Section: Cell Damage From Spargingmentioning

confidence: 99%

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

Hu¹,

2011

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“…Addition of the nonionic block copolymer Pluronic F-68 to the culture medium has been shown to protect mammalian cells (Kilburn and Webb, 1968;Handa et al, 1987;Handa-Corrigan et al, 1989;Passini and Goochee, 1989;Reuveny et al, 1986;Radlett et al, 1971) and insect cells (Murhammer andGoochee, 1988, 1990;Maiorella et al, 1988) from the detrimental effects of sparging. Pluronic F-68, which has a total average molecular weight of 8400, consists of a center block of poly(oxypropy1ene) (20 ?6 by weight) and blocks of poly(oxyethy1ene) at both ends (Stanton, 1957;Swim and Parker, 1960;Schmolka, 1972Schmolka, ,1977. The specific mechanism of Pluronic F-68 protection is unclear.…”

Section: Introductionmentioning

confidence: 99%

Sparged Animal Cell Bioreactors: Mechanism of Cell Damage and Pluronic F‐68 Protection

Murhammer

Goochee

1990

Biotechnology Progress

170

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Pluronic F-68 is a widely used protective agent in sparged animal cell bioreactors. In this study, the attachment-independent Spodoptera frugiperda Sf9 insect cell line was used to explore the mechanism of this protective effect and the nature of cell damage in sparged bioreactors. First, bubble incorporation via cavitation or vortexing was induced by increasing the agitation rate in a surface-aerated bioreactor; insect cells were rapidly killed under these conditions of the absence of polyols. Supplementing the medium with 0.2% (w/v) Pluronic F-68, however, fully protected the cells. Next, cell growth was compared in two airlift bioreactors with similar geometry but different sparger design; one of these bioreactors consisted of a thin membrane distributor, while the other consisted of a porous stainless steel distributor. The flow rates and bubble sizes were comparable in the two bioreactors. Supplementing the medium with 0.2% (w/v) Pluronic F-68 provided full protection to cells growing in the bioreactor with the membrane distributor but provided essentially no protection in the bioreactor with the stainless steel distributor. These results strongly suggest that cell damage can occur in the vicinity of the gas distributor. In addition, these results demonstrate that bubble size and gas flow rate are not the only important considerations of cell damage in sparged bioreactors. A model of cell death in sparged bioreactors is presented.

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“…Despite studying a significant number of compounds, these studies still show that Pluronic F-68 (PF-68), which was one of the first protective additivies advocated, remains the most effective (Kilburn and Webb, 1968;Runyan and Geyer, 1963;Swim and Parker, 1960). However, as was also demonstrated by Ma et al (2004), as the cell density increases, the effectiveness of PF-68 decreases.…”

Section: Introductionmentioning

confidence: 88%

An investigation of small‐molecule surfactants to potentially replace pluronic F‐68 for reducing bubble‐associated cell damage

Rathman

Chalmers

2008

Biotech & Bioengineering

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It is well known that bubble rupture has a detrimental effect on mammalian cells. As a result, Pluronic F-68 (PF-68), a nonionic surfactant, is commonly used to reduce bubble-associated cell damage in sparged bioreactors. While PF-68 is currently effective, there is a concern with respect to its decrease in effectiveness as cell concentrations increase (Ma et al., 2004, Biotechnol Prog 20:1183-1191). In addition, having more than one effective surfactant for cell culture is also highly desirable. Given the empirical nature in which PF-68 was initially discovered as a cell culture additive, a structure-performance study of small molecule surfactants, a distinct group which have been previously investigated for other purposes, was performed in an attempt to find a replacement for PF-68. In this study, a generic platform was established to initially screen both the type and concentration of these surfactants for cytotoxicity. Promising candidates where then evaluated for their ability to rapidly lower the surface tension (dynamic surface tension) of culture media and their ability to prevent cell-bubble attachment in a specially developed bubble creation and collection system. Several promising small- molecule surfactants, and their effective concentration, were identified, which can reduce cell-bubble attachment efficiently without being harmful to cells.

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Effect of Pluronic F68 on Growth of Fibroblasts in Suspension on Rotary Shaker

Cited by 45 publications

References 1 publication

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

Sparged Animal Cell Bioreactors: Mechanism of Cell Damage and Pluronic F‐68 Protection

An investigation of small‐molecule surfactants to potentially replace pluronic F‐68 for reducing bubble‐associated cell damage

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