Single-Cell Technologies to Understand the Mechanisms of Cellular Adaptation in Chemostats

Abstract

There is a growing interest in continuous manufacturing within the bioprocessing community. In this context, the chemostat process is an important unit operation. The current application of chemostat processes in industry is limited although many high yielding processes are reported in literature. In order to reach the full potential of the chemostat in continuous manufacture, the output should be constant. However, adaptation is often observed resulting in changed productivities over time. The observed adaptation can be coupled to the selective pressure of the nutrient-limited environment in the chemostat. We argue that population heterogeneity should be taken into account when studying adaptation in the chemostat. We propose to investigate adaptation at the single-cell level and discuss the potential of different single-cell technologies, which could be used to increase the understanding of the phenomena. Currently, none of the discussed single-cell technologies fulfill all our criteria but in combination they may reveal important information, which can be used to understand and potentially control the adaptation.

Microbioreactors for nutrient‐controlled microbial cultures: Bridging the gap between bioprocess development and industrial use

“…Similar strategies even enable studies on co-cultures of cells in chemostat-like conditions, see Figure 5F. [29] For extensive reviews on applying chamber-based microfluidic devices for studying morphology, heterogeneity, growth, and communication of microorganisms in a high-throughput manner and at single-cell resolution, we refer to Grunberger et al, Burmeister et al and Wright et al [30][31][32]…”

Microbioreactors for nutrient-controlled microbial cultures: Bridging the gap between early bioprocess development and industrial scale use

“…Similar strategies even enable studies on co-cultures of cells in chemostat-like conditions, see Figure.5(f ) [26]. For extensive reviews on applying chamber-based microfluidic devices for studying morphology, heterogeneity, growth, and communication of microorganisms in a high-throughput manner and at single-cell resolution, we refer to [29][30][31].…”

Emergence of Phenotypically Distinct Subpopulations Is a Factor in Adaptation of Recombinant Saccharomyces cerevisiae under Glucose-Limited Conditions

“…The chemostat mode of cultivation is an efficient strategy for industrial production processes, especially in continuous manufacturing, as the cells in the bioreactor can be kept in a constant growth environment for long periods with a constant output (Wright, Rønnest, & Sonnenschein, 2020). In this steady-state growth environment, the cells are exposed to nutrient limitation, e.g.…”