On-line monitoring of dielectrical properties of yeast cells during a stress-model alcoholic fermentation

Tibayrenc, Pierre; Preziosi-Belloy, Laurence; Ghommidh, C.

doi:10.1016/j.procbio.2010.08.007

Cited by 27 publications

(22 citation statements)

References 39 publications

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“…Concerning cell death, f c signal monitoring has already proven its value by detecting cell death of yeast, CHO, and Vero cells 27, 28, 65. For both CHO cells and Saccharomyces yeasts, the increase of f c signal was observed concomitantly with cell death inception and was explained by the augmentation of the intracellular conductivity, σ i .…”

Section: Resultsmentioning

confidence: 99%

“…As described in various studies, the cell dielectric properties of cell membrane capacitance ( C M ) and intracellular conductivity (σ i ) could be expressed in two dielectric measures acquired on‐line, the maximal permittivity (Δε max ) and the characteristic frequency of the cell population ( f c ) 27. Dielectric spectroscopy has been increasingly used to monitor a wide range of bioprocesses, from mammalian cell culture to microbiological systems 28–36. Nevertheless, very few studies have described the application of this technology to viral production processes 33, 37, 38.…”

Section: Introductionmentioning

confidence: 99%

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Real‐time monitoring of influenza virus production kinetics in HEK293 cell cultures

Petiot

Kamen

2012

Biotechnology Progress

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There is an increased interest from the vaccine industry to use mammalian cell cultures for influenza vaccine manufacturing. Therefore, it became important to study the influenza infection mechanism, the viral–host interaction, and the replication kinetics from a bioprocessing stand point to maximize the influenza viral production yield in cell culture. In the present work, influenza replication kinetics was studied in HEK293 cells. Two infection conditions were evaluated, a low (0.01) and a high multiplicity of infection (1.0). Critical time points of the viral production cycle (infection, protein synthesis, viral assembly and budding, viral release, and host‐cell death) were identified in small‐scale cell cultures. Additionally, cell growth, viability, and viral titers were monitored in the viral production process. The infection state of the cultivated cell population was assessed by influenza immunolabeling throughout the culture period. Influenza virus production kinetics were also on‐line monitored by dielectric spectroscopy and successfully correlated to real‐time capacitance measures. Overall, this work provided insights into the mechanisms associated with the infection of human HEK293 cell line by the influenza virus and demonstrated, once again, the usefulness of multifrequency scanning permittivity for in‐line monitoring and supervision of cell‐based viral production processes. Published 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Real‐time monitoring of influenza virus production kinetics in HEK293 cell cultures

Petiot

Kamen

2012

Biotechnology Progress

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“…A sterilizable capacitance probe commercialized as a “biomass monitor” has been available over the last decade. Bulk capacitance and conductance of yeast and mammalian cell cultures can be monitored by a dielectric probe positioned within the bioreactor (Ansorge et al, ; Ducommun et al, ; Knabben et al, ; Opel et al, ; Tibayrenc et al, ). The monitored capacitance of the bulk cell suspension, C , is a function of the effective dielectric permittivity of the suspension, ϵ which is in turn a function of the biovolume.…”

Section: Introductionmentioning

confidence: 99%

The changing dielectric properties of CHO cells can be used to determine early apoptotic events in a bioprocess

Braasch

Nikolic-Jaric

Cabel

et al. 2013

Biotech & Bioengineering

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To ensure maximum productivity of recombinant proteins it is desirable to prolong cell viability during a mammalian cell bioprocess, and therefore important to carefully monitor cell density and viability. In this study, five different and independent methods of monitoring were applied to Chinese hamster ovary (CHO) cells grown in a batch culture in a controlled bioreactor to determine cell density and/or cell viability. They included: a particle counter, trypan blue exclusion (Cedex), an in situ bulk capacitance probe, an off-line fluorescent flow cytometer, and a prototype dielectrophoretic (DEP) cytometer. These various techniques gave similar values during the exponential growth phase. However, beyond the exponential growth phase the viability measurements diverged. Fluorescent flow cytometry with a range of fluorescent markers was used to investigate this divergence and to establish the progress of cell apoptosis: the cell density estimates by the intermediate stage apoptosis assay agreed with those obtained by the bulk capacitance probe and the early stage apoptosis assay viability measurements correlated well with the DEP cytometer. The trypan blue assay showed higher estimates of viable cell density and viability compared to the capacitance probe or the DEP cytometer. The DEP cytometer measures the dielectric properties of individual cells and identified at least two populations of cells, each with a distinct polarizability. As verified by comparison with the Nexin assay, one population was associated with viable (non-apoptotic) cells and the other with apoptotic cells. From the end of the exponential through the stationary and decline stages there was a gradual shift of cell count from the viable into the apoptotic population. However, the two populations maintained their individual dielectric properties throughout this shift. This leads to the conclusion that changes in bulk dielectric properties of cultures might be better modeled as shifts in cells between different dielectric sub-populations, rather than assuming a homogeneous dielectric population. This shows that bulk dielectric probes are sensitive to the early apoptotic changes in cells. DEP cytometry offers a novel and unique technology for analyzing and characterizing mammalian cells based on their dielectric properties, and suggests a potential application of the device as a low-cost, label-free, electronic monitor of physiological changes in cells.

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“…Se han utilizado a lo largo de la historia algunas especies de levaduras del género Saccharomyces y en concreto, Saccharomyces cerevisiae (Figura 4), conocida como levadura de cerveza o levadura del pan debido a su empleo como fermento (Arias, 2012). Es una de las levaduras con mayor impacto económico y social, ya que es usada en las industrias agroalimentarias y en la producción de productos químicos y proteínas heterólogas (Cipollina, Vai, Porro & Hatzis, 2007), así como en la producción de etanol (Tibayrenc, Preziosi-Belloy & Ghommidh, 2011).…”

Section: La Levadura Saccharomyces Cerevisiaeunclassified

Modelización del crecimiento de una población microbiana en medio líquido y su implicación en el estudio de la morfología celular a través del análisis digital de imágenes

Viteri

Blanco

Carbó

et al. 2017

Model. Sci. Educ. Learn.

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Modelización del crecimiento de una población microbiana en medio líquido y su implicación en el estudio de la morfología celular a través del análisis digital de imágenes Modeling the growth of a microbial population in a liquid medium and its involvement of the study of cellular morphology trough digital image analysiś Angel Viteri, Mónica Blanco, Rosa Carbó, Marta Ginovart Universitat Politècnica de Catalunya angel.viteri.leon@gmail.com, monica.blanco@upc.edu, rosa.carbo@upc.edu, marta.ginovart@upc.edu AbstractCuando la levadura crece en un medio líquido rico en nutrientes genera una curva de crecimiento que presenta principalmente tres fases: fase de adaptación (o fase lag), fase exponencial (o fase log), y fase estacionaria. El modelo de Buchanan es un modelo lineal a trozos, simple pero suficientemente bueno para obtener estimaciones de los principales parámetros cinéticos de crecimiento microbiano. La morfometría de las levaduras que se reproducen por gemación (con células madre y células hija) informa sobre el estado de las poblaciones y su capacidad productiva. Se han analizado de forma conjunta resultados experimentales en relación al número de células de S. cerevisiae creciendo en un medio de cultivo líquido cerrado con glucosa y con dos concentraciones diferentes de oxígeno (condiciones aerobias y microaerófilas), y un conjunto de imágenes digitales tomadas de estas células creciendo. Las distribuciones de valores obtenidos para los parámetros morfológicos de las células (área, perímetro, diámetro mayor, diámetro menor, circularidad y elongación) se han relacionado con las distintas fases de crecimiento identificadas con el modelo Buchanan ajustado. La utilización de un modelo de crecimiento poblacional y los parámetros cinéticos estimados junto con el análisis de imágenes digitales de células individuales ha generado una aproximación muy atractiva del sistema, permitiendo conjugar conceptos matemáticos y biológicos a la vez que realizar un estudio a dos niveles distintos del sistema, a nivel poblacional y a nivel individual.When the yeast is grown in a nutrient-rich liquid medium, its growth curve consists of three main phases: adaptation (or lag) phase, exponential (or log) phase and stationary phase. Buchanan's model is a piece-wise linear model which, although being a rather simple model, turns out to be good enough to estimate the main kinetic parameters of microbial growth. The morphometry of yeast cells, which can reproduce by budding (with mother cells and daughter cells), provides information about the state of yeast populations and their productive capacity. Experimental results in connection with the number of cells of S. cerevisiae growing in closed liquid culture with glucose, and under two different oxygen conditions, aerobic and microaerophilic, were jointly analysed with digital images of the growing cells. The observed distributions of the morphological parameters (area, perimeter, major diameter, minor diameter, circularity and elongation) were compared with the growth phases identi...

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On-line monitoring of dielectrical properties of yeast cells during a stress-model alcoholic fermentation

Cited by 27 publications

References 39 publications

Real‐time monitoring of influenza virus production kinetics in HEK293 cell cultures

Real‐time monitoring of influenza virus production kinetics in HEK293 cell cultures

The changing dielectric properties of CHO cells can be used to determine early apoptotic events in a bioprocess

Modelización del crecimiento de una población microbiana en medio líquido y su implicación en el estudio de la morfología celular a través del análisis digital de imágenes

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