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Journal of General Microbiology, 1959
Several methods for growing animal cells in suspension culture were examined, to find the most efficient in terms of cells produced in a given time for the minimum of medium and attention. Continuous medium flow was more efficient than batch-culture, and the preferred system was to add medium to a culture vessel regularly in small doses via a time-switch-controlled solenoid closure a t a rate similar to the growth rate; a turbidimetric safety device ensured that cell density did not drop below levels permitting growth. A mixture of galactose or fructose (6 g./l.) and glucose (2 g./l.) allowed better pH control than glucose alone (6 g./l.). Doubling times of 14-16 hr. were obtained on occasion; the gas-phase oxygen concentration for fastest growth depended on cell density and was frequently less than atmospheric, suggesting that these cells can behave as microaerophils. ' maximal ' medium (medium CSV.6, Table 1) giving fastest growth and highest
Cytotechnology, 1995
A murine hybridoma cell line producing a monoclonal antibody against penicillin-G-amidase and a murine transfectoma cell line secreting a monovalent chimeric human/mouse Fab-antibody fragment were cultivated in three different media (serum-containing, low protein serum-free, and iron-rich protein-free) in flask cultures, stirred reactors and a fixed bed reactor. In static batch cultures in flasks both cell lines showed similar good growth in all three media. In suspension in a stirred reactor, the hybridoma cell line could be cultivated satisfactory only in serum-containing medium. In low protein serum-free medium, Pluronic F68 had to be added to protect the hybridoma cells against shear stress. But even with this supplement only batch, not chemostat mode was possible. In iron-rich protein-free medium the hybridoma cells grew also in continuous chemostat mode, but the stability of the culture was low. The transfectoma cell line did not grow in stirred reactors in any of the three media. Good results with both cell lines were obtained in fixed bed experiments, where the cells were immobilized in macroporous Siran |-carriers. The media, which were optimized in flask cultures, could be used without any further adaptation in the fixed bed reactor. Immobilization improved the stability and reliability of cultures of non-adherent animal cells in serum-free media tremendously compared to suspension cultures in stirred reactors. The volume-specific glucose uptake rate, an indicator of the activity of the immobilized cells, was similar in all three media. Deviations in the metabolism of immobilized and suspended cells seem to be mainly due to low oxygen concentrations within the macroporous carriers, where the cells are supplied with oxygen only by diffusion.
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The culture of animal cells is one of the major aspects of science which serves as a foundation for most of our recent discoveries. The major areas of application include cancer research, vaccine manufacturing, recombinant protein production, drug selection and improvement, gene therapy, stem cell biology, monoclonal antibody production, in vitro fertilization technology, cryopreservation and in vitro production of hormones. Cells can be propagated, expanded and divided into identical replicates, which can be characterized, purified and preserved by freezing. This article reviews the basic aspects of animal cell culture for modern day research.