title: Quantitative and Functional Requirements for Bioluminescent Cancer Models.
abstract: BACKGROUND
  Bioluminescent cancer models are widely used but detailed quantification of the luciferase signal and functional comparison with a non-transfected control cell line are generally lacking. In the present study, we provide quantitative and functional tests for luciferase-transfected cells.
  
  
  METHODS
  We quantified the luciferase expression in BLM and HCT8/E11 transfected cancer cells, and examined the effect of long-term luciferin exposure. The present study also investigated functional differences between parental and transfected cancer cells.
  
  
  RESULTS
  Our results showed that quantification of different single-cell-derived populations are superior with droplet digital polymerase chain reaction. Quantification of luciferase protein level and luciferase bioluminescent activity is only useful when there is a significant difference in copy number. Continuous exposure of cell cultures to luciferin leads to inhibitory effects on mitochondrial activity, cell growth and bioluminescence. These inhibitory effects correlate with luciferase copy number. Cell culture and mouse xenograft assays showed no significant functional differences between luciferase-transfected and parental cells.
  
  
  CONCLUSIONS
  Luciferase-transfected cells should be validated by quantitative and functional assays before starting large-scale experiments.
authors.affiliation: Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University, Ghent, Belgium.
authors.forename: Lynn
authors.initials: L
authors.lastname: Feys
authors.affiliation: iMinds-IBiTech-MEDISIP, Department of Electronics and Information System, Ghent University, Ghent, Belgium.
authors.forename: Benedicte
authors.initials: B
authors.lastname: Descamps
authors.affiliation: iMinds-IBiTech-MEDISIP, Department of Electronics and Information System, Ghent University, Ghent, Belgium.
authors.forename: Christian
authors.initials: C
authors.lastname: Vanhove
authors.affiliation: Department of Biomedical Science, HoGent, Ghent, Belgium.
authors.forename: Stefan
authors.initials: S
authors.lastname: Vermeulen
authors.affiliation: Center of Medical Genetics, Ghent University, Ghent, Belgium.
authors.forename: J O
authors.initials: JO
authors.lastname: Vandesompele
authors.affiliation: Center of Medical Genetics, Ghent University, Ghent, Belgium.
authors.forename: Katrien
authors.initials: K
authors.lastname: Vanderheyden
authors.affiliation: Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
authors.forename: Kathy
authors.initials: K
authors.lastname: Messens
authors.affiliation: Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University, Ghent, Belgium.
authors.forename: Marc
authors.initials: M
authors.lastname: Bracke
authors.affiliation: Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University, Ghent, Belgium Olivier.dewever@ugent.be.
authors.forename: Olivier
authors.initials: O
authors.lastname: De Wever
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D008156:Luciferases / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009369:Neoplasms / Q000378:metabolism*; D014162:Transfection / Q000379:methods
chemical_list: D008156:Luciferases
keywords: BLM; Bioluminescence; cancer model; in vivo imaging; mouse xenograft