On‐line monitoring of cell mass in mammalian cell cultures by acoustic densitometry

Kilburn, Douglas G.; Fitzpatrick, Paula; Blake-Coleman, B.C.; Clarke, David J.; Griffiths, J. B.

doi:10.1002/bit.260331103

Cited by 36 publications

(8 citation statements)

References 2 publications

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“…Besides, direct methods based on viable cell density estimation from physical measurements were also implemented. Thus we could mention the nuclear magnetic resonance spectroscopy (NMR) (Bradamante et al 2004), the acoustic resonance densitometry (ARD) (Kilburn et al 1989), the absorbance or scattering (Card et al 2008), the fluorescence (Teixeira et al 2009), the dielectric spectroscopy or the realtime imaging (Rudolph et al 2008) which were used for this purpose. Only few of these methods are now commercialized, most of them being difficult to set up or presenting a poor sensitivity .…”

Section: Introductionmentioning

confidence: 99%

Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes

Petiot¹,

El-Wajgali²,

Esteban³

et al. 2012

Cytotechnology

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This study proposes an easy to use in situ device, based on multi-frequency permittivity measurements, to monitor the growth and death of attached Vero cells cultivated on microporous microcarriers, without any cell sampling. Vero cell densities were on-line quantified up to 10(6) cell mL(-1). Some parameters which could potentially impact Vero cell morphological and physiological states were assessed through different culture operating conditions, such as media formulation or medium feed-harvest during cell growth phase. A new method of in situ cell death detection with dielectric spectroscopy was also successfully implemented. Thus, through permittivity frequency scanning, major rises of the apoptotic cell population in bioreactor cultures were detected by monitoring the characteristic frequency of the cell population, f(c), which is one of the culture dielectric parameters. Both cell density quantification and cell apoptosis detection are strategic information in cell-based production processes as they are involved in major events of the process, such as scale-up or choice of the viral infection conditions. This new application of dielectric spectroscopy to adherent cell culture processes makes it a very promising tool for risk-mitigation strategy in industrial processes. Therefore, our results contribute to the development of Process Analytical Technology in cell-based industrial processes.

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Section: Introductionmentioning

confidence: 99%

Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes

Petiot¹,

El-Wajgali²,

Esteban³

et al. 2012

Cytotechnology

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“…Considering the importance to CHO cell culture as well as other mammalian cell‐based culture, various PAT tools have been proposed by instrumentation firms to monitor the viable cell density (VCD) in real‐time. These proposed tools include dielectric spectroscopy, near‐infrared spectroscopy, Raman spectroscopy, in situ microscopy, acoustic resonance densitometry, soft‐sensor‐based approaches, and many others. Among these methodologies, dielectric spectroscopy has been claimed to be one of the most reliable to monitor VCD due to its simplicity, robustness and fast signal acquisition.…”

Section: Introductionmentioning

confidence: 99%

Dielectric spectroscopy‐based estimation ofVCDinCHOcell culture

Lee

Carvell

Brorson

et al. 2014

J of Chemical Tech & Biotech

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BACKGROUND In mammalian cell culture processes, viable cell density (VCD) is one of the most crucial parameters for monitoring of the current status of cell growth and protein production. Among the various real‐time monitoring tools available for VCD in Chinese hamster ovary (CHO) cell culture, dielectric spectroscopy is one of the more reliable due to its simplicity, robustness and accuracy. These attributes enable efficient monitoring and control of the corresponding processes. However, one of the major drawbacks of dielectric spectroscopy is frequent deterioration of accuracy during the late‐growth phase of CHO cell culture. This can be caused by abrupt changes in the cell physiology, as well as environmental conditions. RESULTS In this study, various kinds of chemometric tools were employed to understand the spectral dynamics of dielectric spectroscopy during extended CHO cell culture, including different cell growth phases. Then, the performance of different multivariate regression models were compared with conventional Cole‐Cole equation‐based approaches. The models were compared in their ability to estimate VCD in CHO cell culture from dielectric spectra from several batches of bioreactor operations. Multivariate analysis of dielectric spectral datasets clearly revealed the time‐varying evolution of spectral dynamics during the transition between cell growth phases. The best prediction performance from the dielectric spectroscopy was realized using a regression model based on locally‐weighted partial least squares (LWPLS). This estimation model best compensated for spectral dynamics observed during the different cell growth phases. CONCLUSION Dielectric spectroscopy combined with LWPLS allows achievement of optimal and stable estimation of VCD during CHO cell culture, regardless of inherent differences in cell growth phases. © 2014 Society of Chemical Industry

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“…As such, a variety of techniques have been explored for their potential to monitor it on-line. These include optical techniques which work by measuring either turbidity or nephelometry [2,3], fluorescence-based instruments [4], acoustic resonance densitometry (ARD)-based probes [5] and in-line microscopy [6,7]. Indirect techniques which establish a correlation between some observed quantity such as acid/alkali consumption or oxygen uptake rate and the biomass have also been employed [8].…”

Section: Introductionmentioning

confidence: 99%

Use of focussed beam reflectance measurement (FBRM) for monitoring changes in biomass concentration

Whelan

Murphy

Pearson

et al. 2012

Bioprocess Biosyst Eng

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The potential of focussed beam reflectance measurement (FBRM) as a tool to monitor changes in biomass concentration was investigated in a number of biological systems. The measurement technique was applied to two morphologically dissimilar plant cell suspension cultures, Morinda citrifolia and Centaurea calcitrapa, to a filamentous bacteria, Streptomyces natalensis, to high density cultures of Escherichia coli and to a murine Sp2/0 hybridoma suspension cell line, 3-2.19. In all cases, the biomass concentration proved to be correlated with total FBRM counts. The nature of the correlation varied between systems and was influenced by the concentration, nature, size and morphology of the particle under investigation.

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On‐line monitoring of cell mass in mammalian cell cultures by acoustic densitometry

Cited by 36 publications

References 2 publications

Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes

Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes

Dielectric spectroscopy‐based estimation ofVCDinCHOcell culture

Use of focussed beam reflectance measurement (FBRM) for monitoring changes in biomass concentration

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