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.
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
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.
Gamblin, S. J. & Skehel, J. J. J. Biol. Chem. 285, 28403–28409 (2010).
Chambers, B. S., Li, Y., Hodinka, R. L. & Hensley, S. E. J. Virol. 88, 10986–10989 (2014).
Lee, H. K. et al. PLoS ONE 8, e79252 (2013).
Tamura, D. et al. Antimicrob. Agents Chemother. 57, 6141–6146 (2013).
Lin, Y. et al. Influenza Other Respir. Viruses 11, 263–274 (2017).
Li, D. et al. J. Clin. Microbiol. 47, 466–468 (2009).
Oh, D. Y., Barr, I. G., Mosse, J. A. & Laurie, K. L. J. Clin. Microbiol. 46, 2189–2194 (2008).
Mohr, P. G., Deng, Y. M. & McKimm-Breschkin, J. L. Virol. J. 12, 67 (2015).
Lin, Y. P. et al. J. Virol. 84, 6769–6781 (2010).
Zhu, X. et al. J. Virol. 86, 13371–13383 (2012).
Connor, R. J., Kawaoka, Y., Webster, R. G. & Paulson, J. C. Virology 205, 17–23 (1994).
Rogers, G. N. & Paulson, J. C. Virology 127, 361–373 (1983).
Stevens, J. et al. J. Mol. Biol. 355, 1143–1155 (2006).
Lin, S. C., Kappes, M. A., Chen, M. C., Lin, C. C. & Wang, T. T. PLoS ONE 12, e0172299 (2017).
Hatakeyama, S. et al. J. Clin. Microbiol. 43, 4139–4146 (2005).
Matrosovich, M., Matrosovich, T., Carr, J., Roberts, N. A. & Klenk, H. D. J. Virol. 77, 8418–8425 (2003).
van Riel, D. et al. Science 312, 399 (2006).
Shinya, K. et al. Nature 440, 435–436 (2006).
Cong, L. et al. Science 339, 819–823 (2013).
Jinek, M. et al. Science 337, 816–821 (2012).
Han, J. et al. Cell Rep. 23, 596–607 (2018).
Shalem, O. et al. Science 343, 84–87 (2014).
Takashima, S. & Tsuji, S. Trends Glycosci. Glycotechnol. 23, 178–193 (2011).
Chu, V. C. & Whittaker, G. R. Proc. Natl Acad. Sci. USA 101, 18153–18158 (2004).
Hidari, K. I. et al. Biochem. Biophys. Res. Commun. 436, 394–399 (2013).
Shibuya, N. et al. J. Biochem. 106, 1098–1103 (1989).
Chambers, B. S., Parkhouse, K., Ross, T. M., Alby, K. & Hensley, S. E. Cell Rep. 12, 1–6 (2015).
Skowronski, D. M. et al. Euro Surveill. 21, 30112 (2016).
Saito, T. et al. J. Med. Virol. 74, 336–343 (2004).
Lenth, R. V. J. Stat. Softw. 69, 1–33 (2016).
Bates, D., Machler, M., Bolker, B. M. & Walker, S. C. J. Stat. Softw. 67, 1–48 (2015).
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).
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.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Takada, K., Kawakami, C., Fan, S. et al. A humanized MDCK cell line for the efficient isolation and propagation of human influenza viruses. Nat Microbiol 4, 1268–1273 (2019). https://doi.org/10.1038/s41564-019-0433-6
Journal of Biological Chemistry (2021)
Reviews in Medical Virology (2020)
Passage of influenza A/H3N2 viruses in human airway cells removes artefactual variants associated with neuraminidase-mediated binding
Journal of General Virology (2020)