Adaptation of Vero cells to suspension growth for rabies virus production in different serum free media

Rourou, Samia; Zakkour, Meriem Ben; Kallel, Héla

doi:10.1016/j.vaccine.2019.05.092

Cited by 34 publications

(29 citation statements)

References 44 publications

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“…Cell adaptation to suspension growth was not every time as straightforward and successful as previously described (Gallo-Ramirez, Nikolay, Genzel, & Reichl, 2015). A stepwise reduction of percentage of FBS was performed to adapt serum-free culture condition, and it usually takes at least 1-2 months (Lee, Park, & Seo, 2020;Rourou, Ben Zakkour, & Kallel, 2019). More recent studies have indicated that during adaptation, certain cell lines may also change their expression of surface receptors, which then affects production purposes (de Los Milagros Bassani Molinas, Beer, Hesse, Wirth, & Wagner, 2014).…”

Section: Discussionmentioning

confidence: 98%

“…In this study, we successfully adapted 293T adherent cells into 293T suspension cells about a week. Those suspension cells had a stable proliferation, and no need for genetically modification (Humbert et al, 2016), microcarriers (Guy et al, 2013) and antiaggregate solvents (Rourou et al, 2019) to stay uniformly dispersed suspension. After 3 d of cultivation, cell density had reached at least 3.8 times.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Establishing 293T suspension cultrue for lentiviral vector production in a stirred bioreactor

Tang¹,

Gu²,

Han³

et al. 2020

Preprint

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293T suspensions cells were first cultured in a big bubble-stirred bioreactors for large-scale lentivirus vectors (LV) production in the study. Direct domesticated approaches were utilized for 293 suspensions cells. We validated of the ability to LV package and drew a growth curve in shake flask. The results show 293T suspension culture was successful, and also retained the capacity for LV production. After facile optimization of the viral package and cryopreservation conditions, 293T suspension cells were inoculated in a 5.5 L bioreactor. Packaged CAR-CD30 lentivirus yield was close to that from the original shake flask culture. Cultivation for 4 days in the bioreactor enabled production of 1.5±0.3×107 TU/mL coarse LV and the lentiviral infection efficiency was 48.6±2.8% in T Cells. The process has merits of low cost as well as more effective, and expected to be highly competitive compared to other bioreactors.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Establishing 293T suspension cultrue for lentiviral vector production in a stirred bioreactor

Tang¹,

Gu²,

Han³

et al. 2020

Preprint

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“…However, this system was not further exploited. A decade later, new efforts were made and two other lab groups described the adaptation of Vero cells to grow in serum-free suspension cultures ( Rourou et al, 2019 ; Shen et al, 2019 ). Shen et al demonstrated the operation of batch and perfusion bioreactors, including biomass monitoring via capacitance, as well as rVSV production in this system.…”

Section: Suspension Culturesmentioning

confidence: 99%

“…Low cell growth rates with doubling times of more than 40 h and the frequent formation of aggregates remain important issues. Media development will certainly play an important role, highlighted by the fact that no commercial media but only in-house media were able to support Vero suspension cultures ( Rourou et al, 2019 ; Shen et al, 2019 ). For example, cell adhesion is heavily dependent on the concentration of the bivalent cation of calcium in the cell surrounding environment and thus media with reduced concentrations could decrease cell aggregation.…”

Section: Suspension Culturesmentioning

confidence: 99%

Vero cell upstream bioprocess development for the production of viral vectors and vaccines

Kießlich

Kamen

2020

Biotechnology Advances

100

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The Vero cell line is considered the most used continuous cell line for the production of viral vectors and vaccines. Historically, it is the first cell line that was approved by the WHO for the production of human vaccines. Comprehensive experimental data on the production of many viruses using the Vero cell line can be found in the literature. However, the vast majority of these processes is relying on the microcarrier technology. While this system is established for the large-scale manufacturing of viral vaccine, it is still quite complex and labor intensive. Moreover, scale-up remains difficult and is limited by the surface area given by the carriers. To overcome these and other drawbacks and to establish more efficient manufacturing processes, it is a priority to further develop the Vero cell platform by applying novel bioprocess technologies. Especially in times like the current COVID-19 pandemic, advanced and scalable platform technologies could provide more efficient and cost-effective solutions to meet the global vaccine demand. Herein, we review the prevailing literature on Vero cell bioprocess development for the production of viral vectors and vaccines with the aim to assess the recent advances in bioprocess development. We critically underline the need for further research activities and describe bottlenecks to improve the Vero cell platform by taking advantage of recent developments in the cell culture engineering field.

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“…Other successful adaptation strategies have already been described, such as a three-step protocol for the adaptation of mammalian cells (Chinese hamster ovary [CHO] cells) to serum-free media, a synthetic surfactant that increases cell viability in serum-free medium after thawing, adaptation approaches for anchorage-dependent mammalian cells to a suspension serum-free culture condition, adaptation of CHO cells to serum-and proteinfree conditions for recombinant protein production, gradual adaptation of a fish cell line to serum-free conditions in 32 days, and adaptation of Vero cells to suspension culture using different serum-free and chemically defined media (Caron, Biaggio, & Swiech, 2018;González Hernández & Fischer, 2007;Jukić, Bubenik, Pavlović, Tušek, & Srček, 2016;Radošević, Dukić, Andlar, Slivac, & Gaurina Srček, 2016;Rourou, Ben Zakkour, & Kallel, 2019;Sinacore et al, 2000).…”

Section: Background Informationmentioning

confidence: 99%

Adaptation of Mammalian Cells to Chemically Defined Media

2019

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In order to circumvent ethical, technical, and economic drawbacks regarding the use of animal serum in cell culturing, it is possible to adapt mammalian cells to serum‐free media. Nowadays, there are several serum‐free formulations available, including fully animal derived–free and chemically defined media, and different adaptation techniques. This article focuses on the gradual adaptation of a mammalian suspension cell culture to a chemically defined medium. The first step is to transfer the cells cultured in medium supplemented with fetal bovine serum (FBS) to a chemically defined medium of your choice, containing the same amount of FBS. The next steps consist of progressively reducing the amount of FBS, while monitoring cell growth and viability up to the complete elimination of FBS. This protocol has been successfully used to adapt THP‐1 cells to a chemically defined medium with similar maximum specific growth rate as those cultured with FBS. © 2019 by John Wiley & Sons, Inc. Basic Protocol: Gradual adaptation to chemically defined medium Alternate Protocol: Direct adaptation to chemically defined medium Support Protocol 1: Determining maximum specific growth rate of a cell culture Support Protocol 2: Cell freezing and thawing

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Adaptation of Vero cells to suspension growth for rabies virus production in different serum free media

Cited by 34 publications

References 44 publications

Establishing 293T suspension cultrue for lentiviral vector production in a stirred bioreactor

Establishing 293T suspension cultrue for lentiviral vector production in a stirred bioreactor

Vero cell upstream bioprocess development for the production of viral vectors and vaccines

Adaptation of Mammalian Cells to Chemically Defined Media

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