title: A human beta cell line with drug inducible excision of immortalizing transgenes.
abstract: OBJECTIVE
  Access to immortalized human pancreatic beta cell lines that are phenotypically close to genuine adult beta cells, represent a major tool to better understand human beta cell physiology and develop new therapeutics for Diabetes. Here we derived a new conditionally immortalized human beta cell line, EndoC-βH3 in which immortalizing transgene can be efficiently removed by simple addition of tamoxifen.
  
  
  METHODS
  We used lentiviral mediated gene transfer to stably integrate a tamoxifen inducible form of CRE (CRE-ERT2) into the recently developed conditionally immortalized EndoC βH2 line. The resulting EndoC-βH3 line was characterized before and after tamoxifen treatment for cell proliferation, insulin content and insulin secretion.
  
  
  RESULTS
  We showed that EndoC-βH3 expressing CRE-ERT2 can be massively amplified in culture. We established an optimized tamoxifen treatment to efficiently excise the immortalizing transgenes resulting in proliferation arrest. In addition, insulin expression raised by 12 fold and insulin content increased by 23 fold reaching 2 μg of insulin per million cells. Such massive increase was accompanied by enhanced insulin secretion upon glucose stimulation. We further observed that tamoxifen treated cells maintained a stable function for 5 weeks in culture.
  
  
  CONCLUSIONS
  EndoC βH3 cell line represents a powerful tool that allows, using a simple and efficient procedure, the massive production of functional non-proliferative human beta cells. Such cells are close to genuine human beta cells and maintain a stable phenotype for 5 weeks in culture.
authors.affiliation: Institut du cerveau et de la moelle (ICM), Biotechnology & Biotherapy Team, 75013 Paris, France; CNRS UMR7225, 75013 Paris, France; INSERM U1127, 75013 Paris, France; Université Pierre et Marie Curie, 75013 Paris, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Benazra
authors.affiliation: Endocells, Pépinière d'entreprises Institut du Cerveau et de la Moelle, 75007 Paris, France.
authors.forename: Marie-José
authors.initials: MJ
authors.lastname: Lecomte
authors.affiliation: Institut du cerveau et de la moelle (ICM), Biotechnology & Biotherapy Team, 75013 Paris, France; CNRS UMR7225, 75013 Paris, France; INSERM U1127, 75013 Paris, France; Université Pierre et Marie Curie, 75013 Paris, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Colace
authors.affiliation: Paul Langerhans Institute of the Helmholtz Center Munich at the University Hospital and Faculty of Medicine, TU Dresden, 01307 Dresden, Germany; German Center for Diabetes Research (DZD e.V), 85764 Neuherberg, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Müller
authors.affiliation: Endocells, Pépinière d'entreprises Institut du Cerveau et de la Moelle, 75007 Paris, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Machado
authors.affiliation: Endocells, Pépinière d'entreprises Institut du Cerveau et de la Moelle, 75007 Paris, France.
authors.forename: Severine
authors.initials: S
authors.lastname: Pechberty
authors.affiliation: Endocells, Pépinière d'entreprises Institut du Cerveau et de la Moelle, 75007 Paris, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Bricout-Neveu
authors.affiliation: Endocells, Pépinière d'entreprises Institut du Cerveau et de la Moelle, 75007 Paris, France.
authors.forename: Maud
authors.initials: M
authors.lastname: Grenier-Godard
authors.affiliation: Paul Langerhans Institute of the Helmholtz Center Munich at the University Hospital and Faculty of Medicine, TU Dresden, 01307 Dresden, Germany; German Center for Diabetes Research (DZD e.V), 85764 Neuherberg, Germany; Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
authors.forename: Michele
authors.initials: M
authors.lastname: Solimena
authors.affiliation: INSERM, U1016, Institut Cochin, Faculté de Médecine, Université Paris Descartes, Sorbonne Paris Cité, 75014 Paris, France.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Scharfmann
authors.affiliation: Endocells, Pépinière d'entreprises Institut du Cerveau et de la Moelle, 75007 Paris, France.
authors.forename: Paul
authors.initials: P
authors.lastname: Czernichow
authors.affiliation: Institut du cerveau et de la moelle (ICM), Biotechnology & Biotherapy Team, 75013 Paris, France; CNRS UMR7225, 75013 Paris, France; INSERM U1127, 75013 Paris, France; Université Pierre et Marie Curie, 75013 Paris, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Ravassard
keywords: Cell engineering; Conditional immortalization; Human beta cell function; Human pancreatic beta cell line; Tamoxifen inducible CRE