Continuous flow bioreactors with cell retention have been increasingly used for the cultivation of mammalian cells. The potential advantages of such bioreactors are high cell concentrations and volumetric productivities. In many reported cases, these systems have shown fluctuations in cell concentrations of various frequency and magnitude. To analyze the dynamics of the fluctuations, a model-based approach is followed. Simulations showed that large fluctuations in biomass resulted in response to fluctuations in the retention ratio when the system is operated at high dilution rate and high cell retention. The dependence of cell concentration fluctuations on variations in dilution rate and retention ratio was established by a cross-correlation statistical analysis on available experimental data. The slower dynamics and the fluctuation propensity of retention systems suggest that continuous culture without retention is more convenient for kinetic studies. In all likelihood, continuous culture with retention can be stabilized by controlling both the retention ratio and the dilution rate.
The environmental effects on developing somatic embryos should be characterized not only by the growth based on biomass, but also by the morphological properties and size. We have previously developed a discrete classifier to separate developing embryos into distinct morphological classes. In this study, a continuous descriptor using the distributions of magnitude of features representing morphological characteristics and size information was used to describe the developing embryo populations. The identity of the population was examined by comparing either the distributions of all features or key features. The method was applied to characterize the kinetics of carrot embryo populations cultivated in the presence and absence of triiodobenzoic acid(TIBA), an inhibitor of auxin polar transport. Optimal sample size for morphological characterization was determined by the invariance of feature distributions with further increase in sample size. The overall growth and substrate consumption kinetics were only slightly affected by the presence of TIBA. However, the distribution of morphological features was significantly affected. The features showing the highest statistical significance were related to those corresponding to the roughness. The continuous descriptor for characterizing developing embryo population is potentially useful for quality control in large-scale operations.
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