Protein‐free culture of VERO cells: a substrate for replication of human pathogenic viruses

Cinatl, Jindrich; Činátl, Jaroslav; Rabenau, Holger F.; Rapp, Jens; Kornhuber, B.; Doerr, H. W.

doi:10.1006/cbir.1993.1152

Cited by 19 publications

(11 citation statements)

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“…However, a number of continuous cell lines such as MDCK, Vero and BHK-21 already have SFM associated with them and can be expanded in SFM. These include influenza virus production in MDCK (Palache et al 1997;Brands et al 1999;Merten et al 1996); polio virus production in Vero (Cinatl et al 1993;Merten et al 1996) and rabies virus production in BHK-21 or Vero (Perrin et al 1995;Frazatti-Gallina et al 2004). In many of these systems the viral yield was equal if not better than the yield in serum-supplemented media (Merten 2002).…”

Section: Serum Supplementationmentioning

confidence: 99%

The role of recombinant proteins in the development of serum-free media

2006

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Early developments in serum-free media led to a variety of formulations in which components normally provided in serum and required for growth (insulin, transferrin, lipid supplements, trace elements) and poorly defined components (extracts, hydrolysates) were added to defined basal media. These additives were mostly animal-derived. Given recent concerns about TSEs (transmissible spongiform encephalopathies) and other adventitious agents, the drive in media formulations must be towards elimination of animal-origin materials while maintaining cell line productivity. The progress made towards removing animal-derived components and the use of recombinant proteins in serum-free media for mammalian cells is reviewed.

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Section: Serum Supplementationmentioning

confidence: 99%

The role of recombinant proteins in the development of serum-free media

2006

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“…To our knowledge, there are a few reports describing Influenza virus production using MDCK cells in commercially available serum-free medium [21,26-28], while that using Vero cells is described in only one recent report [35] although the medium used contains animal components. Related literature described serum-free media for Vero cells [36,37] and microcarrier bioreactor processes for the production of other viruses using Vero cells [37-46]. It is important to bridge this gap to provide a scalable animal-component free, serum-free platform for researchers and academics to produce different Influenza viruses using Vero cells.…”

Section: Introductionmentioning

confidence: 99%

Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells

Chen

Poh

Dietzsch³

et al. 2011

BMC Biotechnol

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BackgroundInfluenza virus is a major health concern that has huge impacts on the human society, and vaccination remains as one of the most effective ways to mitigate this disease. Comparing the two types of commercially available Influenza vaccine, the live attenuated virus vaccine is more cross-reactive and easier to administer than the traditional inactivated vaccines. One promising live attenuated Influenza vaccine that has completed Phase I clinical trial is deltaFLU, a deletion mutant lacking the viral Nonstructural Protein 1 (NS1) gene. As a consequence of this gene deletion, this mutant virus can only propagate effectively in cells with a deficient interferon-mediated antiviral response. To demonstrate the manufacturability of this vaccine candidate, a batch bioreactor production process using adherent Vero cells on microcarriers in commercially available animal-component free, serum-free media is described.ResultsFive commercially available animal-component free, serum-free media (SFM) were evaluated for growth of Vero cells in agitated Cytodex 1 spinner flask microcarrier cultures. EX-CELL Vero SFM achieved the highest cell concentration of 2.6 × 10^6 cells/ml, whereas other SFM achieved about 1.2 × 10^6 cells/ml. Time points for infection between the late exponential and stationary phases of cell growth had no significant effect in the final virus titres. A virus yield of 7.6 Log10 TCID50/ml was achieved using trypsin concentration of 10 μg/ml and MOI of 0.001. The Influenza vaccine production process was scaled up to a 3 liter controlled stirred tank bioreactor to achieve a cell density of 2.7 × 10^6 cells/ml and virus titre of 8.3 Log10 TCID50/ml. Finally, the bioreactor system was tested for the production of the corresponding wild type H1N1 Influenza virus, which is conventionally used in the production of inactivated vaccine. High virus titres of up to 10 Log10 TCID50/ml were achieved.ConclusionsWe describe for the first time the production of Influenza viruses using Vero cells in commercially available animal-component free, serum-free medium. This work can be used as a basis for efficient production of attenuated as well as wild type Influenza virus for research and vaccine production.

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“…The cell manufacturing process will be more rapid, better controlled and more easily increased for large-scale vaccine production. Growth of virus in serum-free or even proteinfree cell culture [102] signiWcantly minimises the risk for microbial contamination or potential allergic reactions to egg components. InXuenza viruses grown in mammalian cell culture were shown to be more similar to those in human clinical specimens compared with their egg-grown counterparts [103,104].…”

Section: Cell Culture-grown Inxuenza Vaccinementioning

confidence: 99%

The threat of avian influenza A (H5N1). Part IV: development of vaccines

Cinatl

Michaelis

2007

Med Microbiol Immunol

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Among emerging and re-emerging infectious diseases, influenza constitutes one of the major threats to mankind. In this review series epidemiologic, virologic and pathologic concerns raised by infections of humans with avian influenza virus A/H5N1 are discussed. This fourth part focuses on vaccine development. Several phase I clinical studies with vaccines against H5 viruses have demonstrated limited efficacy compared to seasonal influenza vaccines. To induce protective immunity two immunisations with increased amounts of H5N1 vaccine were required. Novel vaccination strategies that are egg- and adjuvant-independent, broadly cross-reactive and long-lasting are highly desirable.

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Protein‐free culture of VERO cells: a substrate for replication of human pathogenic viruses

Cited by 19 publications

References 0 publications

The role of recombinant proteins in the development of serum-free media

The role of recombinant proteins in the development of serum-free media

Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells

The threat of avian influenza A (H5N1). Part IV: development of vaccines

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