Applied Microbiology and Biotechnology

, Volume 73, Issue 6, pp 1423–1434

Hybridoma Ped-2E9 cells cultured under modified conditions can sensitively detect Listeria monocytogenes and Bacillus cereus

  • Pratik Banerjee
  • Mark T. Morgan
  • Jenna L. Rickus
  • Kathy Ragheb
  • Carlos Corvalan
  • J. Paul Robinson
  • Arun K. Bhunia
Applied Microbial and Cell Physiology

DOI: 10.1007/s00253-006-0622-0

Cite this article as:
Banerjee, P., Morgan, M.T., Rickus, J.L. et al. Appl Microbiol Biotechnol (2007) 73: 1423. doi:10.1007/s00253-006-0622-0

Abstract

Lymphocyte origin hybridoma Ped-2E9 cell-based cytotoxicity assay can detect virulent Listeria or Bacillus species, and its application in a cell-based biosensor for onsite use would be very attractive. However, maintaining enough viable cells on a sensor platform for a prolonged duration is a challenging task. In this study, key factors affecting the survival and growth of Ped-2E9 cells under modified conditions were investigated. When the Ped-2E9 cells were grown in media containing 5% fetal bovine serum in sealed tubes without any replenishment of nutrients or exogenous CO2 supply, a large portion of the cells remained viable for 6 to 7 days and cells entered into G0/G1 resting phase. The media pH change was negligible and no cell death was observed in the first 4 days, then cells sequentially underwent apoptotic (fourth day onward) phase until day 7 after which a majority was dead. Subsequent cytotoxicity testing of 3- to 7-day stored Ped-2E9 cells sensitively detected virulent Listeria and Bacillus species. These data strongly suggest that Ped-2E9 cells can be maintained in viable state for 6 days in a sealed tube mimicking the environment in a potential sensor device for onsite use without the need for expensive cell culture facilities.

Keywords

Ped-2E9 Hybridoma Listeria monocytogenes Bacillus Cytotoxicity Cell-based sensor 

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Pratik Banerjee
    • 1
  • Mark T. Morgan
    • 2
  • Jenna L. Rickus
    • 3
  • Kathy Ragheb
    • 4
  • Carlos Corvalan
    • 2
  • J. Paul Robinson
    • 4
  • Arun K. Bhunia
    • 1
  1. 1.Molecular Food Microbiology Laboratory, Department of Food SciencePurdue UniversityWest LafayetteUSA
  2. 2.Department of Food SciencePurdue UniversityWest LafayetteUSA
  3. 3.Department of Agricultural and Biological Engineering and Weldon School of Biomedical EngineeringPurdue UniversityWest LafayetteUSA
  4. 4.Purdue University Cytometry Laboratory, the Bindley Bioscience CenterPurdue UniversityWest LafayetteUSA

Personalised recommendations