title: Hybridoma Ped-2E9 cells cultured under modified conditions can sensitively detect Listeria monocytogenes and Bacillus cereus.
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 CO(2) 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.
authors.affiliation: Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Pratik
authors.initials: P
authors.lastname: Banerjee
authors.forename: Mark T
authors.initials: MT
authors.lastname: Morgan
authors.forename: Jenna L
authors.initials: JL
authors.lastname: Rickus
authors.forename: Kathy
authors.initials: K
authors.lastname: Ragheb
authors.forename: Carlos
authors.initials: C
authors.lastname: Corvalan
authors.forename: J Paul
authors.initials: JP
authors.lastname: Robinson
authors.forename: Arun K
authors.initials: AK
authors.lastname: Bhunia
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001409:Bacillus cereus / Q000254:growth & development / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism / Q000494:pharmacology; D015374:Biosensing Techniques / Q000379:methods*; D002245:Carbon Dioxide / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D006825:Hybridomas / Q000166:cytology* / Q000187:drug effects / Q000382:microbiology; D006863:Hydrogen-Ion Concentration; D008089:Listeria monocytogenes / Q000254:growth & development / Q000378:metabolism*; D051379:Mice; D013997:Time Factors
chemical_list: D001427:Bacterial Toxins; D002245:Carbon Dioxide; D004247:DNA