Death rate in a small air-lift loop reactor of vero cells grown on solid microcarriers and in macroporous microcarriers

Martens, Dirk E.; Nollen, Ellen A. A.; Hardeveld, M.; Groot, C.A.M. van der Velden-de; Gooijer, C.D. de; Beuvery, E. C.; Tramper, J.

doi:10.1007/bf00364836

Cited by 24 publications

(19 citation statements)

References 35 publications

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“…Porous MCs may satisfy the need for a larger surface area for cell attachment, because cells grow outside as well as inside the MC, and for cell protection against the detrimental hydrodynamic forces (Adema et al, 1990;Martens, 1997;Shiragami et al, 1993). On the other hand, the internal cells may be so isolated from the environment that they may be deprived of nutrients and oxygen and spared from virus particle infection.…”

Section: Discussionmentioning

confidence: 97%

“…In spite of good productivity obtained in cell cultures grown on solid MCs (Kennard and Piret, 1995;Mendonça et al, 2002;Montagnon et al, 1981Montagnon et al, , 1984, attempts to achieve larger surface areas led to the development of porous MCs, which can provide higher cell growth surface area compared with solid MCs, due to the attachment and cell growth inside the pores and not only on the peripheral MC area. Porous MCs were shown to allow higher cell/bead loadings, support higher agitation rates, and have the ability to protect the cells against detrimental hydrodynamic forces generated by air sparging or collision of two MCs, or by impact of an MC on the impeller, probes, and wall (Bancel and Hu, 1996;Cahn, 1990;Kennard and Piret, 1995;Lim et al, 1992;Martens et al, 1997;Mignot et al, 1990;Ng et al, 1996;Ohlson et al, 1994, Preissmann et al, 1997Reiter et al, 1990;Shiragami et al, 1993;Zhaolie et al, 1999).…”

Section: Introductionmentioning

confidence: 94%

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Rabies virus production in high vero cell density cultures on macroporous microcarriers

Yokomizo

Antoniazzi

Galdino

et al. 2004

Biotech & Bioengineering

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The purpose of the study was to investigate the rabies virus multiplication in Vero cell cultures performed on porous microcarriers, MCs (cellulose-Cytopore and gelatin-Cultispher G), which provide higher available surface area compared with solid (nonporous) MCs (DEAE-Cytodex 1). In a set of experiments performed at the same MC concentration (MCs per milliliter), cell densities regularly obtained in porous MC cultures were comparable, but almost twice as high as those in solid MC cultures. In addition, 41.1 +/- 3.9-, 35.2 +/- 2-, and 19.6 +/- 5.8-fold increases in cell concentration, relative to the initial cell number, along with maximum rabies virus titers of 6.3 +/- 0.3 x 10(4), 5 +/- 0.1 x 10(4), and 4.3 +/- 0.2 x 10(4) FFD(50)/mL were observed in Cytopore, Cultispher G, and Cytodex 1 MC cultures, respectively. When higher concentrations of MCs were employed, lower performances of virus production and MC-cell occupation (cells per MC or cells per square millimeter) were observed. Cell attachment to MCs was shown to be faster for Cytopore MCs and Cytodex 1 MCs than for Cultispher G MCs. Concerning the kinetics of cell multiplication on MCs, exponential cell growth, at similar specific cell growth rates, took place on Cytopore, Cultispher G, and Cytodex 1 MCs. In addition, cell densities as high as 2.1 +/- 0.2 x 10(6) cells/mL on Cytopore MCs, 1.8 +/- 0.1 x 10(6) cells/mL on Cultispher G MCs, and 1 +/- 0.3 x 10(6) cells/mL on Cytodex 1 MCs were regularly obtained in batch cultures. Optical as well as scanning and transmission electron microscopy studies carried out to analyze MC structure, MC cell occupation, and cell permissivity to virus infection demonstrated that there was uniform cell distribution in the external and internal areas of the MCs, suggesting an efficiency of virus synthesis. Our results indicate the usefulness of these supports for rabies virus antigen production, as well as possibilities for further optimization.

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Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 94%

Rabies virus production in high vero cell density cultures on macroporous microcarriers

Yokomizo

Antoniazzi

Galdino

et al. 2004

Biotech & Bioengineering

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“…The more recent porous containingmicrocarriers which were made from cellulose (16) have an average pore diameter of about 30 µM (17). Thus, the cells in the interior of the macro porous microcarrier are shielded from fluid flow and are hence not susceptible to the detrimental effects of high shear (18). This raises the possibility that potential gradients of nutrients, oxygen, and metabolites along the radius of the microcarrier may cause growth limitation (17).…”

Section: Discussionmentioning

confidence: 99%

Dextran-based Nanocarriers as Efficient Media Delivery Vehicles to Cell Production Bioreactors

Anton¹,

Kambalapally²,

Thomas³

et al. 2010

Nano BioMed ENG

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Decreasing the volume of media required to maintain cell viability not only reduces contamination of bioreactors from the upstream process, but may contribute to cost-containment measures in the biopharmaceutical industry. Based on our recent finding that dextran-containing nanocarriers increased CHO cell density up to 20 fold compared to its cellulose-containing microcarrier counterpart (manuscript submitted), we then investigated the possibility of reducing media volume to maintain cell viability, by utilizing the same dextran-based nanocarrier prepared from a self assembling nanoemulsion (SANE) method, and an adherent Chinese hamster ovary (CHO) cell culture line to evaluate media volume requirements. At the same 60 mL volume of media, cell viability after day 3 was 6 fold greater in CHO cells exposed to dextran-containing nanocarriers compared to cellulose-based microcarriers. When CHO cells were exposed to 60 mL of media containing dextran-based nanocarriers compared to 100 mL of media for cellulose-microcarriers, at day 6, cell density was up to 7 fold greater. Similarly, cell lysate protein concentrations at day 6 was nearly 3 fold greater for CHO cells exposed to dextran-containing nanocarriers compared to the cellulose-based microcarriers. Furthermore, nanocarriers had 59% greater glucose concentration, used as a measure of the polymer dextran and cellulose content levels in the nanocarriers and microcarriers, respectively. In conclusion, nanocarriers with increased numbers of dextran molecules, developed in these studies may be useful to further optimize media volume requirements for maximum culture growth.

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“…The linear viral DNA method has been found to work for either the polyhedrin or pl0 loci of AcMN-PV, provided a cognate transfer vector is used (Kitts et al, 1990;Kitts, 1995;Martens et al, 1995). AcM-NPV viral DNA linearized at either the polyhedrin or p10 locus is commercially available from Invitrogen or PharMingen.…”

Section: Recombination Between a Transfer Vector And Linear Viral Dnamentioning

confidence: 99%

Insect Cell Culture: Fundamental and Applied Aspects

Vlak¹,

Gooijer²,

Tramper³

et al. 2002

Current Applications of Cell Culture Engineering

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Death rate in a small air-lift loop reactor of vero cells grown on solid microcarriers and in macroporous microcarriers

Cited by 24 publications

References 35 publications

Rabies virus production in high vero cell density cultures on macroporous microcarriers

Rabies virus production in high vero cell density cultures on macroporous microcarriers

Dextran-based Nanocarriers as Efficient Media Delivery Vehicles to Cell Production Bioreactors

Insect Cell Culture: Fundamental and Applied Aspects

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