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Anoikis-resistant MDCK cells carrying susceptibilities to TNF-α and verotoxin that are suitable for influenza virus cultivation

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Abstract

Madin-Darby canine kidney (MDCK) cells were originally anchorage-dependent epithelial cells. Here, we have isolated a novel MDCK-derived cell population, termed 6 M-4, by means of culturing MDCK cells in suspension for nearly 6 months in the presence of Streptomyces griseus metalloendopeptidase (MEP). The isolated cells showed unique proliferation characteristics, which differed from parental MDCK cells. They proliferated adherently on a polystyrene matrix, but proliferated non-adherently both in the presence of MEP and on a non-adhesive matrix coated with poly 2-methacryloyloxyethyl phosphorylcholine (MPC). The 6 M-4 cells consisted of at least two cell types. One type, termed 6 M-4-TR7, would not grow in soft agar and showed a novel phenotype in that the cells were susceptible to both TNF-α and verotoxin 1 (VT1). In addition, the isolated adhesion-independent cells sustained epithelial traits of parental MDCK cells. We further show that these MDCK-derivative cells are suitable for influenza virus cultivation. Hemagglutination (HA) titers of influenzaviruses A and B were increased in the suspension culture of 6 M-4-TR7 cells supplemented with the MEP in comparison to adherently growing cells in the presence of trypsin.

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Abbreviations

MEM:

minimum essential medium

MEP:

Streptomyces griseus metalloendopeptidase

MPC:

poly 2-methacryloyloxyethyl phosphorylcholine

rTNF-α:

recombinant tumor necrosis factor α

VT1:

verotoxin 1

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Acknowledgements

This study was supported by “High-Tech Research Center” Project for Private Universities: matching fund subsidy from the Ministry of Education, Culture, Sports, Science and Technology of Japan, 2002–2006. We greatly appreciate Dr. K. Tsutsumi for his discussion and critical reading of the manuscript. We would like to thank Mrs. Yaegashi for excellent technical assistance in cell culture.

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Authors and Affiliations

  1. Department of Microbiology, School of Medicine, Iwate Medical University, 19-1 Uchimaru, Morioka, Iwate, 020-8505, Japan

    Reiko Tsutsumi & Shigehiro Sato

  2. Institute of Enzyme Research, Osaka, Japan

    Shigemi Fujisaki

  3. Department of Radiology, School of Dentistry, Iwate Medical University, Uchimaru, Morioka, Iwate, 020-8505, Japan

    Masanori Shozushima

  4. Research Institute for Environmental Sciences and Public Health of Iwate, 4 Iioka-shinden, Morioka, Iwate, 020-0852, Japan

    Koichi Saito

Authors
  1. Reiko Tsutsumi
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  2. Shigemi Fujisaki
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  3. Masanori Shozushima
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  4. Koichi Saito
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  5. Shigehiro Sato
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Correspondence to Shigehiro Sato.

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Tsutsumi, R., Fujisaki, S., Shozushima, M. et al. Anoikis-resistant MDCK cells carrying susceptibilities to TNF-α and verotoxin that are suitable for influenza virus cultivation. Cytotechnology 52, 71–85 (2006). https://doi.org/10.1007/s10616-006-9032-5

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  • DOI: https://doi.org/10.1007/s10616-006-9032-5

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