Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes
Identifieur interne : 000973 ( Hal/Curation ); précédent : 000972; suivant : 000974Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes
Auteurs : Emma Petiot [France] ; Amal El-Wajgali [France] ; Geoffrey Esteban [France] ; Cecile Geny [France] ; Hervé Pinton [France] ; Annie Marc [France]Source :
- Cytotechnology [ 0920-9069 ] ; 2012.
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Abstract
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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DOI: 10.1007/s10616-011-9421-2
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<profileDesc><textClass><keywords scheme="mix" xml:lang="en"><term>Adherent Vero cells</term>
<term>Apoptosis</term>
<term>BIOLOGICAL CELLS</term>
<term>CULTURE PROCESSES</term>
<term>Cell density</term>
<term>DIELECTRIC-SPECTROSCOPY</term>
<term>HL-60 CELLS</term>
<term>INTRACELLULAR CONDUCTIVITY</term>
<term>In situ monitoring</term>
<term>MEMBRANE CAPACITANCE</term>
<term>Multi-frequency permittivity</term>
<term>ONLINE DETERMINATION</term>
<term>PERMITTIVITY MEASUREMENTS</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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<hal api="V3"><titleStmt><title xml:lang="en">Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes</title>
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<surname>Petiot</surname>
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<email>Emma.Petiot@cnrc-nrc.gc.ca</email>
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<author role="aut"><persName><forename type="first">Amal</forename>
<surname>El-Wajgali</surname>
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<author role="aut"><persName><forename type="first">Geoffrey</forename>
<surname>Esteban</surname>
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<author role="aut"><persName><forename type="first">Cecile</forename>
<surname>Geny</surname>
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<author role="aut"><persName><forename type="first">Hervé</forename>
<surname>Pinton</surname>
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<idno type="halauthor">807032</idno>
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<author role="aut"><persName><forename type="first">Annie</forename>
<surname>Marc</surname>
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<email>annie.marc@univ-lorraine.fr</email>
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<editor role="depositor"><persName><forename>Gabriel</forename>
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<email>gabriel.wild@univ-lorraine.fr</email>
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<editionStmt><edition n="v1" type="current"><date type="whenSubmitted">2013-01-17 18:01:48</date>
<date type="whenWritten">2012</date>
<date type="whenModified">2016-05-10 16:53:03</date>
<date type="whenReleased">2013-01-17 18:01:48</date>
<date type="whenProduced">2012</date>
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<idno type="halRefHtml">Cytotechnology, Springer Verlag, 2012, 64 (4), pp.429-441. <10.1007/s10616-011-9421-2></idno>
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<textClass><keywords scheme="author"><term xml:lang="en">Adherent Vero cells</term>
<term xml:lang="en">In situ monitoring</term>
<term xml:lang="en">Multi-frequency permittivity</term>
<term xml:lang="en">Cell density</term>
<term xml:lang="en">Apoptosis</term>
<term xml:lang="en">SERUM-FREE MEDIUM</term>
<term xml:lang="en">DIELECTRIC-SPECTROSCOPY</term>
<term xml:lang="en">PERMITTIVITY MEASUREMENTS</term>
<term xml:lang="en">MEMBRANE CAPACITANCE</term>
<term xml:lang="en">INTRACELLULAR CONDUCTIVITY</term>
<term xml:lang="en">ONLINE DETERMINATION</term>
<term xml:lang="en">CULTURE PROCESSES</term>
<term xml:lang="en">BIOLOGICAL CELLS</term>
<term xml:lang="en">HL-60 CELLS</term>
<term xml:lang="en">VIRUS</term>
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<abstract xml:lang="en">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.</abstract>
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