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A humanized MDCK cell line for the efficient isolation and propagation of human influenza viruses

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Abstract

Here, we developed hCK, a Madin-Darby canine kidney (MDCK) cell line that expresses high levels of human influenza virus receptors and low levels of avian virus receptors. hCK cells supported human A/H3N2 influenza virus isolation and growth much more effectively than conventional MDCK or human virus receptor-overexpressing (AX4) cells. A/H3N2 viruses propagated in hCK cells also maintained higher genetic stability than those propagated in MDCK and AX4 cells.

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Data availability

The data that support the findings of this study are available from the corresponding authors on request. DNA sequencing data from this study are available under the NCBI BioProject accession number PRJNA525907.

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Acknowledgements

We thank S. Watson for scientific editing. We also thank N. Midorikawa for technical assistance. In addition, we thank E. Adachi, T. Koibuchi, T. Kikuchi, M. Koga, H. Yotsuyanagi, A. Tokita and N. Wada for providing us with the clinical specimens from patients with influenza-like symptoms. Flow cytometry was performed in the IMSUT FACS Core laboratory; we acknowledge the IMSUT FACS Core laboratory for assistance with the flow cytometric analysis. This research was supported by Leading Advanced Projects for medical innovation (LEAP) from the Japan Agency for Medical Research and Development (AMED) (JP18am001007), by Grants-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Science, Sports, and Technology (MEXT) of Japan (nos. 16H06429, 16K21723 and 16H06434), by the Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) from AMED (JP18fm0108006), by an e-ASIA Joint Research Program from AMED (JP17jm0210042), by a Research Program on Emerging and Re-emerging Infectious Diseases from AMED (JP18fk0108104), by a Grant-in-Aid for JSPS Research Fellows (17J04123) and by the NIAID-funded Center for Research on Influenza Pathogenesis (CRIP; HHSN272201400008C).

Author information

K.T., C.K., S.F., S.C., G.Z., C.G., K.S., S.T., Y.S.-T., J.D., Z.K., D.K., H.v.B., S.Y. and M.I. performed the experiments. K.T., C.K., S.F., S.T., T.J.S.L., T.W., M.I. and Y.K. planned the experiments and/or analysed the data. K.T., T.J.S.L., M.I. and Y.K. wrote the manuscript.

Competing interests

K.T., C.K., S.F., S.C., G.Z., C.G., K.S., S.T., T.J.S.L., J.D., Z.K., D.K., H.v.B., Y.S.-T., S.Y., T.W. and M.I. declare no competing interests. Y.K. has received speaker’s honoraria from Toyama Chemical and Astellas and grant support from Chugai Pharmaceuticals, Daiichi Sankyo Pharmaceutical, Toyama Chemical, Tauns Laboratories and Otsuka Pharmaceutical, and is a founder of FluGen.

Correspondence to Masaki Imai or Yoshihiro Kawaoka.

Supplementary information

  1. Supplementary Information

    Supplementary Data and Discussion, Supplementary Methods, Supplementary Figures 1–3, Supplementary Tables 1–8 and Supplementary References.

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Fig. 1: Generation and characterization of hCK cells and their sensitivity to human influenza virus growth and isolation.