Density inhibition and growth kinetics of anchorage-dependent cells
Abstract
To understand the phenomenon of density-dependent inhibition is not only essential for understanding the growth control in anchorage-dependent cells but also necessary for quantitative study of their growth kinetics. Previous studies in this area were all based on the results from batch cultures, which may be biased by the depletion of essential nutrients or the accumulation of cell secretions. A flow system was therefore designed to eliminate the variations in chemical environment during the growth. By comparing the results obtained from batch cultures with those from the flow system, it was found that the secretion of growth factors significantly affect the growth rate of Chinese Hamster Ovary (CHO) cells in a batch culture. With the help of time-lapse cinematography and image analysis, we found that limitation of mass transfer rate may be responsible for the density-dependent inhibition of CHO cells. Based on our observations, a stochastic model was developed to simulate the effect of density inhibition of CHO cells on a flat surface and the surface of a spherical particle. A satisfactory agreement between model simulations and experimental data was obtained. According to the observations and analyses of CHO cells grown on two different surfaces at various serum concentrations, mass transfer may also contribute to the phenomenon of anchorage dependence of CHO cells.
Degree
Ph.D.
Advisors
Tsao, Purdue University.
Subject Area
Chemical engineering|Cellular biology
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