High‐Density culture of mouse–human hybridoma in serum‐free defined medium

Takazawa, Yoshiharu; Tokashiki, Michiyuki; Murakami, Hiroki; Yamada, Koji; Ômura, Hirohisa

doi:10.1002/bit.260310210

Cited by 44 publications

(11 citation statements)

References 4 publications

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“…This result is consistent with the previous ones in which a lower plasma membrane fluidity was correlated with lower shear sensitivity. Takasawa et al 49 have found that addition of very low density lipoproteins (VLDL) to high density cultures of hybridomas can minimize cell damage caused by agitation. It is well known that VLDL readily transfer cholesterol into the plasma membrane of mammalian cells.48 Hence, it can be argued that the protective effect described by Takasawa et al 49 was possibly due to an increased cholesterol content in the cells which effectively decreased the PMF.…”

Section: Effect Of Cholesterolmentioning

confidence: 99%

The role of the plasma membrane fluidity on the shear sensitivity of hybridomas grown under hydrodynamic stress

Ramı́rez¹,

Mutharasan²

1990

Biotech & Bioengineering

120

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The role of the plasma membrane fluidity (PMF) on the shear sensitivity of HB-32 hybridomas to laminar fluid shear was investigated. Steady-state fluorescence anisotropy (r(s)) of the cationic fluorescent probe 1-[4-(trimethylamino) phenyl]-6-phenylhexa-1,3,5-triene, was used to evaluate the PMF of whole hybridoma cells. The PMF was manipulated by the addition of the anesthetic benzyl alcohol, by temperature changes and by cholesterol enrichment. The effect of these PMF modifying procedures on the shear sensitivity of HB-32 was assessed by exposing the cells to defined levels of laminar shear stress in a Couette flow device. Conditions that resulted in lower r(s) values (indicating higher PMF) yielded a more fragile cell. Batch cultivations supplemented with the shear protective agent Pluronic(R) F-68 showed higher values of r(s) compared to control experiments during various growth phases, suggesting that the protective mechanism of Pluronic F-68 relies on its ability to decrease the PMF through direct interaction with the plasma membrane. The protective mechanism of serum against turbulent fluid shear is also discussed from analysis of growth and death kinetics of agitated and static cultures at increasing serum levels. The results of this study show that the fluid state of the plasma membrane is important in determining the integrity of hybridomas when exposed to lethal shear levels. It is concluded that increasing membrane fluidity correlates with increasing shear sensitivity.

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Section: Effect Of Cholesterolmentioning

confidence: 99%

The role of the plasma membrane fluidity on the shear sensitivity of hybridomas grown under hydrodynamic stress

Ramı́rez¹,

Mutharasan²

1990

Biotech & Bioengineering

120

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“…Also worthy of note here is that significant process improvement is most likely with perfusion type systems based on recent published work (e.g. Takazawa et al, 1988;Klerx et al, 1988). Table 6 summarises a few of the considerations to be made when evaluating cell culture systems for in vitro production of antibodies.…”

Section: Discussionmentioning

confidence: 99%

“…more ancillary equipment such as pumps, holding tanks and continuous centrifuges and thus more susceptible to contamination and a possible reduction of antibody specificity at low dilution rates (Harbour et al, 1988). Perfusion systems appear widely in the literature (e.g., Tharakan and Chau, 1986;Takazawa et al, 1988;Altshuler et al, 1986;Klerx et al, 1988;Reuveny et al, 1985Reuveny et al, , 1987Butler et al, 1983). Despite their obvious advantages there are problems associated with the high populations of non-viable cells due to mass transfer problems and operational problems such as sterility maintenance, arising during long production runs.…”

Section: Discussionmentioning

confidence: 99%

A comparison of different culture methods for hybridoma propagation and monoclonal antibody production

et al. 1990

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A major variable to consider in the production of biologicals from mammalian cell cultures is the mode of operation, be it a batch, continuous, perfusion, fed-batch or other production method. The final choice must consider a number of fundamental and economic issues. Here we present some antibody production data from different cell lines using different modes of production and discuss the important factors for consideration in choosing a production strategy. It was found that the productivity of batch cultures was lower than that obtained in continuous and perfused cultures, but that productivity could be improved by implementing suitable feeding strategies. The antibody productivity of one cell line, MCL1, during exponential phase was not affected by media type or glucose level. The maximum productivity of two cell lines in continuous culture was found to occur at dilution rates below the maximum, from 0.019 to 0.030 hr-1.

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“…Therefore, each charge can have a different ability for promoting cell growth and product formation. The use of a totally defined serum free medium can overcome this problem if it is possible to adapt the cells to this conditions without loss of productivity (Takazawa et al, 1988).…”

Section: Introductionmentioning

confidence: 99%

Optimization of serum-free fermentation processes for antibody production

1991

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Serum free fermentation procedures of cell cultures have got a wide application in production of biochemicals. But, cells cultured in serum free media in general are more sensitive to changes in culture condition, especially to nutrient limitation. There are no substances from serum which can support the cells when conditions are changing. In this study special attention is directed to amino acid utilization of mouse hybridoma in batch, chemostat and perfusion fermentations. Detailed data are presented which show the considerable difference of amino acid consumption rates in different fermentation modes. Already, in batch mode there are differences of the two investigated mouse hybridoma cell lines, although they are derived from the same myeloma line. In chemostat running at a dilution rate representing maximal growth rate most of the consumption rates are significant higher than in batch. On the other hand, in perfusion mode the rates are lower than in batch. This indicates clearly the different conditions of the fermentation modes. Therefore, it is necessary to develop serum free processes under the desired production conditions. An accurate anaylsis of the process is strongly recommended.

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High‐Density culture of mouse–human hybridoma in serum‐free defined medium

Cited by 44 publications

References 4 publications

The role of the plasma membrane fluidity on the shear sensitivity of hybridomas grown under hydrodynamic stress

The role of the plasma membrane fluidity on the shear sensitivity of hybridomas grown under hydrodynamic stress

A comparison of different culture methods for hybridoma propagation and monoclonal antibody production

Optimization of serum-free fermentation processes for antibody production

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