LAPSE:2021.0054
Published Article
LAPSE:2021.0054
Propagation and Molecular Characterization of Fowl Adenovirus Serotype 8b Isolates in Chicken Embryo Liver Cells Adapted on Cytodex™ 1 Microcarrier Using Stirred Tank Bioreactor
Chidozie C. Ugwu, Mohd Hair-Bejo, Mat I. Nurulfiza, Abdul R. Omar, Aini Ideris
February 22, 2021
Large volume production of vaccine virus is essential for prevention and control of viral diseases. The objectives of this study were to propagate Fowl adenovirus (FAdV) isolate (UPM08136) in chicken embryo liver (CEL) cells adapted to Cytodex™ 1 microcarriers using stirred tank bioreactor (STB) and molecularly characterize the virus. CEL cells were prepared and seeded onto prepared Cytodex™ 1 microcarriers and incubated first in stationary phase for 3 h and in STB at 37 °C, 5% CO2, and 20 rpm for 24 h. The CEL cells were infected with FAdV isolate (UPM08136) passage 5 (UPM08136CELP5) or passage 20 (UPM08136CELP20) and monitored until cell detachment. Immunofluorescence, TCID50, sequencing, alignment of hexon and fiber genes, and phylogenetic analysis were carried out. CEL cells were adapted well to Cytodex™ 1 microcarriers and successfully propagated the FAdV isolates in STB with virus titer of 107.5 (UPM08136CELP5B1) and 106.5 (UPM08136CELP20B1) TCID50/mL. These isolates clustered with the reference FAdV serotype 8b in the same evolutionary clade. The molecular characteristics remained unchanged, except for a point substitution at position 4 of the hexon gene of UPM08136CELP20B1, suggesting that propagation of the FAdV isolate in STB is stable and suitable for large volume production and could be a breakthrough in the scale-up process.
Keywords
bioreactor, chicken liver cell, Cytodex 1™ microcarrier, fowl adenovirus 8b, PCR
Subject
Suggested Citation
Ugwu CC, Hair-Bejo M, Nurulfiza MI, Omar AR, Ideris A. Propagation and Molecular Characterization of Fowl Adenovirus Serotype 8b Isolates in Chicken Embryo Liver Cells Adapted on Cytodex™ 1 Microcarrier Using Stirred Tank Bioreactor. (2021). LAPSE:2021.0054
Author Affiliations
Ugwu CC: Faculty of Veterinary Medicine, University Putra Malaysia, Serdang 43400, Selangor, Malaysia; Department of Animal Science and Technology, Federal University of Technology, Owerri 460114, Imo State, Nigeria
Hair-Bejo M: Faculty of Veterinary Medicine, University Putra Malaysia, Serdang 43400, Selangor, Malaysia; Institute of Bioscience, University Putra Malaysia, Serdang 43400, Selangor, Malaysia
Nurulfiza MI: Institute of Bioscience, University Putra Malaysia, Serdang 43400, Selangor, Malaysia; Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, Serdang 43400, Selangor, Malaysia
Omar AR: Faculty of Veterinary Medicine, University Putra Malaysia, Serdang 43400, Selangor, Malaysia; Institute of Bioscience, University Putra Malaysia, Serdang 43400, Selangor, Malaysia [ORCID]
Ideris A: Faculty of Veterinary Medicine, University Putra Malaysia, Serdang 43400, Selangor, Malaysia; Institute of Bioscience, University Putra Malaysia, Serdang 43400, Selangor, Malaysia
Journal Name
Processes
Volume
8
Issue
9
Article Number
E1065
Year
2020
Publication Date
2020-09-01
Published Version
ISSN
2227-9717
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Original Submission
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PII: pr8091065, Publication Type: Journal Article
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LAPSE:2021.0054
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doi:10.3390/pr8091065
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Feb 22, 2021
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Feb 22, 2021
 
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Calvin Tsay
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