|SHITTU, ISMAILA - Consultant|
|ZHU, ZIYING - University Of Georgia|
|STICE, STEVEN - University Of Georgia|
|WEST, FRANKLIN - University Of Georgia|
|DONADEU, MERITXELL - Galvmed|
|DUNGU, BAPTISTE - Galvmed|
Submitted to: Biologicals
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/5/2015
Publication Date: 11/14/2015
Publication URL: https://handle.nal.usda.gov/10113/5537799
Citation: Shittu, I., Zhu, Z., Stice, S.L., West, F.D., Donadeu, M., Dungu, B., Fadly, A.M., Afonso, C.L. 2015. Development, characterization, and optimization of a new suspension chicken-induced pluripotent stem cell line for the production of Newcastle disease vaccine. Biologicals. 44(1)24-32 doi: 10.1016/j.biologicals.2015.09.002.
Interpretive Summary: Newcastle disease virus (NDV) and avian influenza are among other very important diseases of poultry that require the use of vaccine viruses that are normally grown in specific pathogen free eggs from chickens. Lack of availability of eggs is often a limiting factor to scale up production of vaccines, and quite often it is a problem in Africa where poultry vaccines are produced by government laboratories. A rapidly growing chicken cell line with good industrial will facilitate production of avian vaccines. Here we have developed a pluripotent cell line that supports the growth of Newcastle disease viruses at high titers while maintaining highly desirable industrial features such as absence of endogenous viruses, high replication rate, growth in suspension, and growth in serum free medium.
Technical Abstract: Traditionally, substrates for production of vaccines have been embryonated eggs or adherent cell culture. The daunting challenge of scaling up these technologies in the face of an outbreak has been a limitation for industrial applicability. Suspension cell lines are better suited in many ways to embryonated eggs due to their ability to grow to high densities and easy scalability to meet urgent demands. In this study, we assess the ability of a newly developed suspension chicken-induced pluripotent stem cell line to support viral replication by determining its capacity to support growth of Newcastle disease virus (NDV) LaSota vaccine strain. The cell characteristics and growth profile, as well as NDV replication, was investigated in cell culture. We showed that cells could grow to a high density of up to 7.0 × 107 cells/mL and showed rapid proliferation with doubling time of 21 hours. Upon infection, a high virus titer of 1.02 × 108 EID50/mL was obtained at 24hrs post infection using a multiplicity of infection (MOI) of 5. One of the major limitations of establishing new cell lines, is the presence of congenitally transmitted as well as endogenous viruses, making them unsuitable for vaccine production. We assessed potential contamination of the cell line for Fowl adenovirus 1, reticuloendotheliosis virus (REV), and chicken infectious anemia by direct polymerase chain reaction (PCR) or reverse transcription (RT)-PCR and found them to be negative. In addition, the cell line was shown to be free of endogenous and exogenous avian leukosis viruses and (REV) by virus isolation tests using both enzyme-linked immuno assay (ELISA) and PCR. In conclusion, this new avian cell line is potentially an excellent candidate for vaccine production due to its highly desirable industrially friendly characteristics.