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Suspension culture process for H9N2 avian influenza virus (strain Re-2)

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Abstract

H9N2 avian influenza virus has caused huge economic loss for the Chinese poultry industry since it was first identified. Vaccination is frequently used as a control method for the disease. Meanwhile suspension culture has become an important tool for the development of influenza vaccines. To optimize the suspension culture conditions for the avian influenza H9N2 virus (Re-2 strain) in Madin-Darby Canine Kidney (MDCK) cells, we studied the culture conditions for cell growth and proliferation parameters for H9N2 virus replication. MDCK cells were successfully cultured in suspension, from a small scale to industrial levels of production, with passage time and initial cell density being optimized. The influence of pH on the culture process in the reactor has been discussed and the process parameters for industrial production were explored via amplification of the 650L reactor. Subsequently, we cultivated cells at high cell density and harvested high amounts of virus, reaching 10log2 (1:1024). Furthermore an animal experiment was conducted to detect antibody. Compared to the chicken embryo virus vaccine, virus cultured from MDCK suspension cells can produce a higher amount of antibodies. The suspension culture process is simple and cost efficient, thus providing a solid foundation for the realization of large-scale avian influenza vaccine production.

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Acknowledgements

We are grateful of Mr Zhaoyang Li, president of Shandong Sinder Technology Co., Ltd for experiment conditions and financially support. We also thanks the technical support and experimental help by the fellow Xiangxia Meng, Yuancang Zhu, Fumeng Wang and Zhiliang Liu.

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Authors and Affiliations

  1. Shandong Sinder Technology Co., Ltd., Xinsheng Street 263, Zhucheng, 262204, Shandong, China

    Honglin Wang & Guicai Song

  2. Department of Biology, Longcheng Middle School, Shunwang Street 204, Zhucheng, 262226, Shandong, China

    Suying Guo

  3. Sinovet (Beijing) Biotechnology Co., Ltd., Kaituo Road 5, Haidian District, Beijing, 100085, China

    Zhenguang Li

  4. Jiangyan Animal Health Inspection Institute, Jiangguan Road 251, Taizhou, 225529, Jiangsu, China

    Xiaoqin Xu

  5. Shandong Zhucheng Animal Husbandry and Veterinary Administration, Dongwu Street 123, Zhucheng, 262200, Shandong, China

    Zexiang Shao

Authors
  1. Honglin Wang
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  2. Suying Guo
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  3. Zhenguang Li
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  4. Xiaoqin Xu
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  6. Guicai Song
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Contributions

HLW and ZGL participated in the design of the study, performed the experiments and drafted the manuscript. SYG and ZXS helped with the cell culture operation. XQX performed the statistical analysis. GCS conceived the study and participated in its design and coordination. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Guicai Song.

Ethics declarations

All experimental procedures have been reviewed and approved by local Animal Care and Use Committee of Shandong Sinder Technology Co., Ltd.

Conflict of interest

The authors declare that they have no conflict of interests.

Funding

The study was financially supported by Shandong Sinder Technology Co., Ltd.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individual participants included in the study.

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Wang, H., Guo, S., Li, Z. et al. Suspension culture process for H9N2 avian influenza virus (strain Re-2). Arch Virol 162, 3051–3059 (2017). https://doi.org/10.1007/s00705-017-3460-8

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  • DOI: https://doi.org/10.1007/s00705-017-3460-8

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