Recent studies have suggested that Bcl-2 can affect cell cycle re-entry by inhibiting the transition from G 0 /G 1 to S phase. In this study, we have taken a novel route to the study of the relationship between Bcl-2 expression and cell cycle progression. Continuous cultures of pEF (control) and Bcl
Determinants and rate laws of growth and death of hybridoma cells in continuous culture
โ Scribed by An-Ping Zeng; Wolf-Dieter Deckwer; Wei-Shou Hu
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 210 KB
- Volume
- 57
- Category
- Article
- ISSN
- 0006-3592
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โฆ Synopsis
Experimental data from six hybridoma cell lines grown under diverse experimental conditions in both normal continuous and perfusion cultures are analyzed with respect to the significance of nutrients and products in determining the growth and death rates of cells and with respect to their mathematical modeling. It is shown that neither nutrients (glucose and glutamine) nor the common products lactic acid, ammonia, and monoclonal antibody can be generally assumed to be the clear-limiting or inhibiting factors for most of the cultures. Correspondingly, none of the unstructured models existing in the literature can be generally applied to describe the experimental data obtained over a relatively wide range of cultivation conditions as considered in this work. Surprisingly, for all cultures the specific growth rate (ยต) almost linearly correlates with the ratio of the viable cell concentration (N V ) to the dilution (perfusion) rate (D). Similarly, the specific death rate (k d ) is a function of the ratio of the total cell concentration (N t ) to the dilution (perfusion) rate. These results strongly suggest the formation of not yet identified critical factors or autoinhibitors that determine both the growth and death rates of hybridoma cells. Based on these observations, simple kinetic models are developed for ยต and k d which describe the experimental data satisfactorily. Analysis of the experimental data with the kinetic models reveals that under the current cultivation conditions the formation rate of the autoinhibitor(s) or the sensitivity of cell growth and death to the autoinhibitor(s) is mainly affected by the medium composition. Irrespective of the cell lines, cells grown on serum-containing media have almost the same model parameters, which are distinctively different from those of cells grown on serum-free media. Furthermore, in contrast to the prevailing view, k d is shown to positively correlate with ยต if the effects of cell concentration and dilution (perfusion) rate are considered. Several important implications of these findings are discussed for the optimization and control of animal cell culture.
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The cell growth and monoclonal antibody production kinetics of hybridoma cell cultures continuously exposed to growth factors and the cognate antigen were investigated. The growth factors were the epidermal growth factor, fibroblast growth factor, and interleukin-2, whereas the antigen was the trini