Strain Agnostic Influenza Virus Propagation in a Serum‐Free, Suspension‐Adapted MDCK Cell Line
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2026-03
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Abstract
<jats:title>ABSTRACT</jats:title> <jats:sec> <jats:title>Background</jats:title> <jats:p>The continuing circulation and evolution of seasonal influenza viruses remains a public health and socioeconomic threat on a global scale. Viral surveillance and vaccination of the public have been relied upon to confer and boost immunity in the population. Traditionally, influenza strains are propagated in embryonated chicken eggs, but this process remains imperfect and subject to genetic drift of the virus and a reliable source of eggs. Cell culture‐based propagation of influenza virus has recently been commercialized, but this method has been difficult to adapt to lab settings.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>Madin‐Darby canine kidney (MDCK) cells were adapted to thrive in serum‐free growth in suspension. The suspension MDCK (sMDCK) line was characterized by measuring replication and viability during routine passage and infection. Fifteen different influenza strains were propagated using this model and were assayed to determine hemagglutination and plaque forming units and compared to influenza strains grown in adherent cell culture. Microneutralization tests were also conducted to ensure each strain maintained the proper antigenicity.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>The cell line was successfully adapted to serum‐free growth in suspension. For each virus strain, the sMDCK platform successfully produced a virus stock in 1–3 days. Additionally, sMDCK progeny virus maintained its antigenicity based on neutralization assays.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusions</jats:title> <jats:p>This simple, scalable method was used to reliably propagate 15 influenza strains with the elimination of costly reagents and animal serum. The results are comparable to traditional methods, and the protocol presented in this work could be adapted to nearly any laboratory setting.</jats:p> </jats:sec>
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Huskey, Jessica B, Michelle L Rock, Pooja V Chaudhary, Emily C Hill, Madeline E Hoover, Nicole M Rideout, Kamerin D Dean, Thomas Scott Alderman, et al. (2026). Strain Agnostic Influenza Virus Propagation in a Serum‐Free, Suspension‐Adapted MDCK Cell Line. Influenza and Other Respiratory Viruses, 20(3). 10.1111/irv.70237 Retrieved from https://hdl.handle.net/10161/34318.
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Scott Alderman
Michael Anthony Moody
Tony Moody, MD is a Professor in the Department of Pediatrics, Division of Infectious Diseases and Professor in the Department of Integrative Immunobiology at Duke University Medical Center. Research in the Moody lab is focused on understanding the B cell responses during infection, vaccination, and disease. The lab has become a resource for human phenotyping, flow characterization, staining and analysis at the Duke Human Vaccine Institute (DHVI). The Moody lab is currently funded to study influenza, syphilis, HIV-1, and emerging infectious diseases.
Dr. Moody is the director of the Duke CIVICs Vaccine Center (DCVC) at (DHVI) and co-director of the Centers for Research of Emerging Infectious Disease Coordinating Center (CREID-CC). Dr. Moody is mPI of a U01 program to develop a syphilis vaccine; this program is a collaboration with mPI Dr. Justin Radolf at the University of Connecticut. Dr. Moody is also the director of the DHVI Accessioning Unit, a biorepository that provides support for work occurring at DHVI and with its many collaborators around the world by providing processing, shipping, and inventory support for a wide array of projects.
Dr. Moody and his team are involved in many networks studying vaccine response including the Collaborative Influenza Vaccine Innovation Centers (CIVICs) and the COVID-19 Prevention Network (CoVPN).
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