From a discrete to continuous actuation for improved real-time control of gene expression in mammalian cells

Postiglione, Lorena; Santorelli, Marco; Tumaini, Barbara; Bernardo, Diego di

doi:10.1016/j.ifacol.2016.12.096

Cited by 6 publications

(6 citation statements)

References 7 publications

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“…The simplest realization of DCE can be done by hydrostatic pressure (Figure 3A, Setup). [ 50,56 ] For the experimental implementation two syringe drivers are used with different media or different concentrations of the same medium. A computer‐controlled stepper motor can control the input transfers of different environments by varying the height between the two syringes or the syringes are controlled electrically (Figure 3A, Principle).…”

Section: Microfluidic Setups For Dynamically Controlled Environments mentioning

confidence: 99%

“…Therefore, this system offers an on/off regulation for the control of the inflow in the microfluidic chip. [ 56 ] This setup is simple and easy to handle (Figure 3A, Specifications). The limitation of the setup is that only low‐frequency modulations in the range of minutes can be created.…”

Section: Microfluidic Setups For Dynamically Controlled Environments mentioning

confidence: 99%

“…Reproduced with permission. [ 50,56 ] Copyright 2019, American Chemical Society. B) Application from Atencia et al [51] illustrates microfluidic chip design with two inlets and pressure pump for DCE between 3 and 12 h. The representative result illustrates dynamic toxicity of Vero cells by cycloheximide pulses.…”

Section: Applicationsmentioning

confidence: 99%

“…This way, they also investigated the gene expression of tetracycline inducible promotor of CHO and could demonstrated a set‐point regulation of gene expression in a proof of concept study. [ 56,77 ]…”

Section: Applicationsmentioning

confidence: 99%

“…Furthermore, automated programmable microfluidic platforms that combine image‐based single‐cell analysis and DCE with feedback control are emerging. [ 50,56,78–80 ] Here, cellular behavior is controlled by real time image analysis of cells, coupled to dynamic environmental control. [ 96,97 ] As soon as cellular threshold values are reached defined medium pulses are triggered (referred as negative feedback control).…”

Section: Technical and Biological Perspectivementioning

confidence: 99%

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Dynamic Environmental Control in Microfluidic Single‐Cell Cultivations: From Concepts to Applications

Täuber

Lieres

Grünberger

2020

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Microfluidic single‐cell cultivation (MSCC) is an emerging field within fundamental as well as applied biology. During the last years, most MSCCs were performed at constant environmental conditions. Recently, MSCC at oscillating and dynamic environmental conditions has started to gain significant interest in the research community for the investigation of cellular behavior. Herein, an overview of this topic is given and microfluidic concepts that enable oscillating and dynamic control of environmental conditions with a focus on medium conditions are discussed, and their application in single‐cell research for the cultivation of both mammalian and microbial cell systems is demonstrated. Furthermore, perspectives for performing MSCC at complex dynamic environmental profiles of single parameters and multiparameters (e.g., pH and O2) in amplitude and time are discussed. The technical progress in this field provides completely new experimental approaches and lays the foundation for systematic analysis of cellular metabolism at fluctuating environments.

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Section: Microfluidic Setups For Dynamically Controlled Environments mentioning

confidence: 99%

Section: Microfluidic Setups For Dynamically Controlled Environments mentioning

confidence: 99%

Section: Applicationsmentioning

confidence: 99%

Section: Applicationsmentioning

confidence: 99%

Section: Technical and Biological Perspectivementioning

confidence: 99%

See 3 more Smart Citations

Dynamic Environmental Control in Microfluidic Single‐Cell Cultivations: From Concepts to Applications

Täuber

Lieres

Grünberger

2020

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Development and application of a cultivation platform for mammalian suspension cell lines with single‐cell resolution

Schmitz

Täuber

Westerwalbesloh

et al. 2020

Biotech & Bioengineering

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In bioproduction processes, cellular heterogeneity can cause unpredictable process outcomes or even provoke process failure. Still, cellular heterogeneity is not examined systematically in bioprocess research and development. One reason for this shortcoming is the applied average bulk analyses, which are not able to detect cell‐to‐cell differences. In this study, we present a microfluidic tool for mammalian single‐cell cultivation (MaSC) of suspension cells. The design of our platform allows cultivation in highly controllable environments. As a model system, Chinese hamster ovary cells (CHO‐K1) were cultivated over 150 h. Growth behavior was analyzed on a single‐cell level and resulted in growth rates between 0.85 and 1.16 day−1. At the same time, heterogeneous growth and division behavior, for example, unequal division time, as well as rare cellular events like polynucleation or reversed mitosis were observed, which would have remained undetected in a standard population analysis based on average measurements. Therefore, MaSC will open the door for systematic single‐cell analysis of mammalian suspension cells. Possible fields of application represent basic research topics like cell‐to‐cell heterogeneity, clonal stability, pharmaceutical drug screening, and stem cell research, as well as bioprocess related topics such as media development and novel scale‐down approaches.

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TFEB dynamical model reveals a novel feedback loop biological mechanism

et al. 2019

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From a discrete to continuous actuation for improved real-time control of gene expression in mammalian cells

Cited by 6 publications

References 7 publications

Dynamic Environmental Control in Microfluidic Single‐Cell Cultivations: From Concepts to Applications

Dynamic Environmental Control in Microfluidic Single‐Cell Cultivations: From Concepts to Applications

Development and application of a cultivation platform for mammalian suspension cell lines with single‐cell resolution

TFEB dynamical model reveals a novel feedback loop biological mechanism

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