Suspension culture of Vero cells for the production of adenovirus type 5

Lee, Deuk-Ki; Park, Jihye; Seo, Dong-Won

doi:10.7774/cevr.2020.9.1.48

Cited by 15 publications

(15 citation statements)

References 13 publications

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“…Using Vero cells that mimic the entry cells of adenoviruses, we managed to replicate HAdV-36 in them, as was done for HAdV-5 7 . Furthermore, we obtained longterm infected cultures by infecting Vero cells with a low MOI (0.5).…”

Section: Resultsmentioning

confidence: 99%

Long-term infection passaging of Human Adenovirus 36 in monkey kidney cells

Alarcon-Valdes

Sanchez-Aguillon

Martínez‐Hernández

et al. 2022

Rev. Inst. Med. trop. S. Paulo

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Human Adenovirus 36 (HAdV-36) has been related to diverse effects on metabolism and may attenuate the lipid accumulation in kidneys with increased adiposity. Some of these effects would be related to viral persistence. However, until now, a model of persistent in vitro infection by HAdV-36 is unknown. In this study, we examined the cells of the Vero lineage to explore their permissiveness to long-term HAdV-36 infection. HAdV-36 was productively replicated in Vero cells and maintained long-term infection for up to 35 cell passages. A subculture was obtained from the cells that survived the primary infection at a low MOI (0.5). The production of the extracellular infectious virus with titers ranging from 10 4 to 10 6 TCID 50 /mL and DNA-bearing cells was detected. In long-term infected cells, the intracellular distribution of viral antigen was demonstrated by performing immunolocalization (IFI) and expression of cell-viral antigen in 50% of cells by flow cytometry, using anti-HAdV-36 hyperimmune rabbit serum. Furthermore, E1a and E4orf1 genes in long-term infected passages showed a decreasing trend. Our preliminary results reveal that renal epithelial monkey cells are permissive for the productive infection of HAdV-36. Vero cell culture long-term infection might be a promising model for addressing the fundamental aspects of the HAdV-36 biology that cannot reveal broadly-used cultures, which do not maintain longterm infection in primary or transformed cells.

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Section: Resultsmentioning

confidence: 99%

Long-term infection passaging of Human Adenovirus 36 in monkey kidney cells

Alarcon-Valdes

Sanchez-Aguillon

Martínez‐Hernández

et al. 2022

Rev. Inst. Med. trop. S. Paulo

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“…In the case of human viruses, in addition to the wide variety of isolated cells derived from this species, many viruses are produced in nonhuman primate cells (Kiesslich and Kamen, 2020). Obviously, cells isolated from other animals allow the replication of human viruses (Tuffereau and Flamand, 1983;Lee et al, 2020a). The capability of a virus to infect a distinct group of cells in the host is defined as tropism, which is often associated with a variety of cellular devices available on the surface of the host cell (Chappell and Dermody, 2015).…”

Section: Host and Cellular Tropismsmentioning

confidence: 99%

Choosing a cellular model to study SARS-CoV-2

Souza

Bideau

Boschi

et al. 2022

Front. Cell. Infect. Microbiol.

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As new pathogens emerge, new challenges must be faced. This is no different in infectious disease research, where identifying the best tools available in laboratories to conduct an investigation can, at least initially, be particularly complicated. However, in the context of an emerging virus, such as SARS-CoV-2, which was recently detected in China and has become a global threat to healthcare systems, developing models of infection and pathogenesis is urgently required. Cell-based approaches are crucial to understanding coronavirus infection biology, growth kinetics, and tropism. Usually, laboratory cell lines are the first line in experimental models to study viral pathogenicity and perform assays aimed at screening antiviral compounds which are efficient at blocking the replication of emerging viruses, saving time and resources, reducing the use of experimental animals. However, determining the ideal cell type can be challenging, especially when several researchers have to adapt their studies to specific requirements. This review strives to guide scientists who are venturing into studying SARS-CoV-2 and help them choose the right cellular models. It revisits basic concepts of virology and presents the currently available in vitro models, their advantages and disadvantages, and the known consequences of each choice.

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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%

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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Suspension culture of Vero cells for the production of adenovirus type 5

Cited by 15 publications

References 13 publications

Long-term infection passaging of Human Adenovirus 36 in monkey kidney cells

Long-term infection passaging of Human Adenovirus 36 in monkey kidney cells

Choosing a cellular model to study SARS-CoV-2

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

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