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Volume 14
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10.3390/vaccines14010052
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Engineering AQP1-Deficient DF-1 Suspension Cells for High-Yield IBDV Production and Vaccine Scale-Up

by
Bingmei Dong
1,
Ruonan Wang
2,
Yu Guan
2,
Xiubao Zhao
2,
Ronghua Li
1,
Qingqing Xu
2,
Hui Li
2,
Qingfang Gao
3,
Shengjie Yao
1,
Shuyu Song
1,
Ashenafi Kiros Wubshet
2,4 and
Na Tang
2,*
1
College of Life Science, Cangzhou Normal University, Guofeng South Avenue 16#, Cangzhou 061001, China
2
Shandong Binzhou Institute of Animal Husbandry and Veterinary Sciences, Binzhou 256600, China
3
Shandong Lvdu Biological Technology Co. Ltd., Binzhou 256600, China
4
College of Veterinary and Animal Sciences, Mekelle University, Mekelle 2084, Ethiopia
*
Author to whom correspondence should be addressed.
Vaccines 2026, 14(1), 52; https://doi.org/10.3390/vaccines14010052
Submission received: 30 November 2025 / Revised: 26 December 2025 / Accepted: 26 December 2025 / Published: 31 December 2025
(This article belongs to the Special Issue Vaccines Against Poultry Viruses)
Review Reports Versions Notes

Abstract

Background: Large-scale production of poultry viral vaccines increasingly requires robust suspension cell platforms. However, most avian cell lines, including DF-1, are strictly anchorage-dependent, limiting scalability. Aquaporin-1 (AQP1) regulates cell–cell adhesion and membrane dynamics, making it a potential target for engineering suspension growth. This study aimed to generate a stable DF-1 suspension cell line via AQP1 disruption and evaluate its potential for enhanced infectious bursal disease virus (IBDV) production. Methodology: DF-1 cells were engineered using a CRISPR/Cas9 ribonucleoprotein system to create a truncated AQP1 gene. DF-1/AQP1− cells were assessed for morphology, tumorigenicity in nude mice, and genetic stability across 20 passages. Suspension growth, cell density, and viability were measured. Cells were infected with IBDV strain BJQ902, and viral titers were compared with wild-type DF-1 and monolayer DF-1/AQP1− cells. Results: DF-1/AQP1− cells maintained normal morphology, were non-tumorigenic, and retained stable AQP1 mutations. They grew as true suspension cultures without adaptation, reaching 4.0 × 106 cells/mL with >95% viability. Suspension DF-1/AQP1− cells cells produced significantly higher viral titers (9.0 log TCID50/mL; 8.63 log EID50/mL) than both monolayer DF-1/AQP1− and wild-type DF-1 cells. Virus production time was shortened in suspension cultures. Conclusions: Targeted AQP1 disruption converts DF-1 cells into a stable, non-tumorigenic suspension cell line with markedly enhanced IBDV production, providing a scalable platform for next-generation avian vaccine manufacturing.
Keywords: DF-1 cells; suspension-adapted culture; aquaporin-1 (AQP1); infectious bursal disease virus (IBDV) DF-1 cells; suspension-adapted culture; aquaporin-1 (AQP1); infectious bursal disease virus (IBDV)

Share and Cite

MDPI and ACS Style

Dong, B.; Wang, R.; Guan, Y.; Zhao, X.; Li, R.; Xu, Q.; Li, H.; Gao, Q.; Yao, S.; Song, S.; et al. Engineering AQP1-Deficient DF-1 Suspension Cells for High-Yield IBDV Production and Vaccine Scale-Up. Vaccines 2026, 14, 52. https://doi.org/10.3390/vaccines14010052

AMA Style

Dong B, Wang R, Guan Y, Zhao X, Li R, Xu Q, Li H, Gao Q, Yao S, Song S, et al. Engineering AQP1-Deficient DF-1 Suspension Cells for High-Yield IBDV Production and Vaccine Scale-Up. Vaccines. 2026; 14(1):52. https://doi.org/10.3390/vaccines14010052

Chicago/Turabian Style

Dong, Bingmei, Ruonan Wang, Yu Guan, Xiubao Zhao, Ronghua Li, Qingqing Xu, Hui Li, Qingfang Gao, Shengjie Yao, Shuyu Song, and et al. 2026. "Engineering AQP1-Deficient DF-1 Suspension Cells for High-Yield IBDV Production and Vaccine Scale-Up" Vaccines 14, no. 1: 52. https://doi.org/10.3390/vaccines14010052

APA Style

Dong, B., Wang, R., Guan, Y., Zhao, X., Li, R., Xu, Q., Li, H., Gao, Q., Yao, S., Song, S., Wubshet, A. K., & Tang, N. (2026). Engineering AQP1-Deficient DF-1 Suspension Cells for High-Yield IBDV Production and Vaccine Scale-Up. Vaccines, 14(1), 52. https://doi.org/10.3390/vaccines14010052

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