Cytotechnology

, Volume 52, Issue 2, pp 71–85 | Cite as

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

  • Reiko Tsutsumi
  • Shigemi Fujisaki
  • Masanori Shozushima
  • Koichi Saito
  • Shigehiro Sato
Original Paper

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.

Keywords

MDCK Anoikis-resistance Cell adherence TNF-α Verotoxin Influenza virus 

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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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Reiko Tsutsumi
    • 1
  • Shigemi Fujisaki
    • 2
  • Masanori Shozushima
    • 3
  • Koichi Saito
    • 4
  • Shigehiro Sato
    • 1
  1. 1.Department of Microbiology, School of MedicineIwate Medical UniversityIwateJapan
  2. 2.Institute of Enzyme ResearchOsakaJapan
  3. 3.Department of Radiology, School of DentistryIwate Medical UniversityIwateJapan
  4. 4.Research Institute for Environmental Sciences and Public Health of IwateIwateJapan

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