Propagation of viruses infecting waterfowl on continuous cell lines of Muscovy duck (<i>Cairina moschata</i>) origin

Mészáros, István; Tóth, Renáta; Bálint, Ádám; Dán, Ádám; Jordan, Ingo; Zádori, Zoltán

doi:10.1080/03079457.2014.939941

Cited by 14 publications

(12 citation statements)

References 32 publications

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“…The growth rate of propagated respective viruses (GPV, DHAV and NDV) in the GEE cell line was started to increase gradually until reached to highest rate at 96, 48 and 72 hpi., where the viral genome replication reached a 10 7.313 , 10 9.774 , and 10 8.633 copies/ml, respectively. In agreement with a previous study [ 7 ] which also showed that the copy number of the propagated GPV and DHAV in muscovy duck fibroblast cell lines peaked with 2x10 6 and 10 6 , respectively. Our finding that the GEE cell line can be supported replication of the GPV was not surprising, as previous data have shown that other cell lines can also be facilitated propagation of the GPV [ 7 ].…”

Section: Discussionsupporting

confidence: 93%

“…In agreement with a previous study [ 7 ] which also showed that the copy number of the propagated GPV and DHAV in muscovy duck fibroblast cell lines peaked with 2x10 6 and 10 6 , respectively. Our finding that the GEE cell line can be supported replication of the GPV was not surprising, as previous data have shown that other cell lines can also be facilitated propagation of the GPV [ 7 ]. However, we were surprised by the ability of the established GEE cell line to assist replication of other avian viruses, such as DHAV and NDV with high infectivity titers compared to other cell lines [ 6 , 8 , 10 , 14 ].…”

Section: Discussionsupporting

confidence: 93%

“…In addition, TCID 50 and ELD 50 assay were used to determine kinetics of infective viral titers of each virus separately at different time points of the infection. It has been reported previously that the GPV can be propagated in goose embryo fibroblasts [ 9 ] and muscovy duck embryo fibroblast cell lines [ 7 ]. To relate our findings to previous studies, the established GEE cell line was evaluated for GPV-infection susceptibility and showed a strong susceptibility to GPV.…”

Section: Discussionmentioning

confidence: 99%

“…Susceptibility of the established cell line to DHAV infection was detected and our results showed that the GEE cell line can efficiently support DHAV virus replication with a maximum virus titer of 10 10.5 TCID 50 /ml at 48 hpi and 10 10.2 ELD 50 /1 ml, which higher than the ELD 50 of propagated DHAV in the embryonated eggs. We and others, have previously shown that Duck epithelial cell line (DEE) [ 14 ], Duck embryo [ 6 ] and Muscovy duck embryo fibroblast cell lines [ 7 ] have been used to facilitate isolation and propagation of the DHAV. While these studies have been demonstrated that the titer of the DHAV was a relatively low compared with the titer of the propagated DHAV in the GEE cell line.…”

Section: Discussionmentioning

confidence: 99%

“…The infected cells were harvested at the indicated time points (12, 24, 36, 48, 72, 96, 108, and 120 hpi) and analyzed using TCID 50 assay as mentioned above and qPCR. The qPCR was done as described earlier [ 7 ]. Briefly, RNA was extracted at different time points (12, 24, 36, 48, 72, 96, 108, and 120 hpi) using RNA Extraction kit (Bioteke).…”

Section: Methodsmentioning

confidence: 99%

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Establishment and evaluation of the goose embryo epithelial (GEE) cell line as a new model for propagation of avian viruses

et al. 2018

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In this study, we report the establishment and characterization of a new epithelial cell line, goose embryonated epithelial cell line (GEE), derived from embryonic goose tissue. The purified GEE cell line can efficiently grow over 65 passages in the M199 medium supplemented with 10% fetal bovine serum at 37°C. Immunofluorescence assay was used to identify purified GEE cells as epithelial cell line by detecting expression of the Keratin-18 and -19. Further characterizations demonstrated that the GEE cell line can be continuously subcultured with (i) a high capacity to replicate for over 65 passages, (ii) a spontaneous epithelial-like morphology, (iii) constant chromosomal features and (iv) without an evidence of converting to tumorigenic cells either in vitro or in vivo study. Moreover, the GEE cell line can be effectively transfected with plasmids expressing reporter genes of different avian viruses, such as VP3, VP1 and F of goose parvo virus (GPV), duck hepatitis virus (DHV), and Newcastle disease virus (NDV), respectively. Finally, the established GEE cell line was evaluated for avian viruses infection susceptibility. Our results showed that the tested GPV, DHAV and NDV were capable to replicate in the new cell line with titers a comparatively higher to the ones detected in the traditional culture system. Accordingly, our established GEE cell line is apparently a suitable in vitro model for transgenic, and infection manipulation studies.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Establishment and evaluation of the goose embryo epithelial (GEE) cell line as a new model for propagation of avian viruses

et al. 2018

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Chicken Infectious Anemia and Circovirus Infections in Commercial Flocks

Schat¹,

Santen²

2019

Diseases of Poultry

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Designing cell lines for viral vaccine production: Where do we stand?

Genzel

2015

Biotechnology Journal

101

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Established animal cells, such as Vero, Madin Darby canine kidney (MDCK) or chicken embryo fibroblasts (CEFs), are still the main cell lines used for viral vaccine production, although new "designer cells" have been available for some years. These designer cell lines were specifically developed as a cell substrate for one application and are well characterized. Later screening for other possible applications widened the product range. These cells grow in suspension in chemically defined media under controlled conditions and can be used for up to 100 passages. Scale-up is easier and current process options allow cultivation in disposable bioreactors at cell concentrations higher than 1 × 10(7) cells/mL. This review covers the limitations of established cell lines and discusses the requirements and screening options for new host cells. Currently available designer cells for viral vaccine production (PER.C6, CAP, AGE1.CR, EB66 cells), together with other new cell lines (PBS-1, QOR/2E11, SogE, MFF-8C1 cells) that were recently described as possible cell substrates are presented. Using current process knowledge and cell line development tools, future upstream processing could resemble today's Chinese hamster ovary (CHO) cell processes for monoclonal antibody production: small scale bioreactors (disposable) in perfusion or fed-batch mode with cell concentrations above 1 × 10(8) cells/mL.

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Propagation of viruses infecting waterfowl on continuous cell lines of Muscovy duck (Cairina moschata) origin

Cited by 14 publications

References 32 publications

Establishment and evaluation of the goose embryo epithelial (GEE) cell line as a new model for propagation of avian viruses

Establishment and evaluation of the goose embryo epithelial (GEE) cell line as a new model for propagation of avian viruses

Chicken Infectious Anemia and Circovirus Infections in Commercial Flocks

Designing cell lines for viral vaccine production: Where do we stand?

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