Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Alerts
  • Advertising/recruitment
  • Subscribe
  • Contact
  • Current Issue
  • Past Issues
  • By specialty
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All...
  • Videos
    • Conversations with Giants in Medicine
    • Author's Takes
  • Reviews
    • View all reviews...
    • Mechanisms Underlying the Metabolic Syndrome (Oct 2019)
    • Reparative Immunology (Jul 2019)
    • Allergy (Apr 2019)
    • Biology of familial cancer predisposition syndromes (Feb 2019)
    • Mitochondrial dysfunction in disease (Aug 2018)
    • Lipid mediators of disease (Jul 2018)
    • Cellular senescence in human disease (Apr 2018)
    • View all review series...
  • Collections
    • Recently published
    • In-Press Preview
    • Commentaries
    • Concise Communication
    • Editorials
    • Viewpoint
    • Scientific Show Stoppers
    • Top read articles
  • Clinical Medicine
  • JCI This Month
    • Current issue
    • Past issues

  • About
  • Editors
  • Consulting Editors
  • For authors
  • Current issue
  • Past issues
  • By specialty
  • Subscribe
  • Alerts
  • Advertise
  • Contact
  • Conversations with Giants in Medicine
  • Author's Takes
  • Recently published
  • Brief Reports
  • Technical Advances
  • Commentaries
  • Editorials
  • Hindsight
  • Review series
  • Reviews
  • The Attending Physician
  • First Author Perspectives
  • Scientific Show Stoppers
  • Top read articles
  • Concise Communication
Development of a conditionally immortalized human pancreatic β cell line
Raphaël Scharfmann, … , Paul Czernichow, Philippe Ravassard
Raphaël Scharfmann, … , Paul Czernichow, Philippe Ravassard
Published May 1, 2014; First published March 25, 2014
Citation Information: J Clin Invest. 2014;124(5):2087-2098. https://doi.org/10.1172/JCI72674.
View: Text | PDF
Categories: Technical Advance Endocrinology

Development of a conditionally immortalized human pancreatic β cell line

  • Text
  • PDF
Abstract

Diabetic patients exhibit a reduction in β cells, which secrete insulin to help regulate glucose homeostasis; however, little is known about the factors that regulate proliferation of these cells in human pancreas. Access to primary human β cells is limited and a challenge for both functional studies and drug discovery progress. We previously reported the generation of a human β cell line (EndoC-βH1) that was generated from human fetal pancreas by targeted oncogenesis followed by in vivo cell differentiation in mice. EndoC-βH1 cells display many functional properties of adult β cells, including expression of β cell markers and insulin secretion following glucose stimulation; however, unlike primary β cells, EndoC-βH1 cells continuously proliferate. Here, we devised a strategy to generate conditionally immortalized human β cell lines based on Cre-mediated excision of the immortalizing transgenes. The resulting cell line (EndoC-βH2) could be massively amplified in vitro. After expansion, transgenes were efficiently excised upon Cre expression, leading to an arrest of cell proliferation and pronounced enhancement of β cell–specific features such as insulin expression, content, and secretion. Our data indicate that excised EndoC-βH2 cells are highly representative of human β cells and should be a valuable tool for further analysis of human β cells.

Authors

Raphaël Scharfmann, Severine Pechberty, Yasmine Hazhouz, Manon von Bülow, Emilie Bricout-Neveu, Maud Grenier-Godard, Fanny Guez, Latif Rachdi, Matthias Lohmann, Paul Czernichow, Philippe Ravassard

×

Figure 1

Strategy to produce reversely immortalized cell line and immunofluorescence analysis of EndoC-βH2 cells.

Options: View larger image (or click on image) Download as PowerPoint
Strategy to produce reversely immortalized cell line and immunofluoresce...
(A) Schematic representation of excisable lentiviral vector used to produce the EndoC-βH2 reversely immortalized cell line. (B) Immunofluorescence analysis of EndoC-βH2 cells. EndoC-βH2 cells stained positive for insulin (INS) and coexpressed C-peptide (C-PEPT), CHGA, PDX1, NKX6.1, SV40 LT, and Ki67. The cells rarely express somatostatin (SST) and do not express glucagon (GCG), amylase (AMY), carboxipeptidase-A (CPA), or SOX9. Nuclei were stained with Hoechst 33342 fluorescent stain (blue). Photographs of all insulin stainings were taken using a 700 ms acquisition time to obtain an unsaturated signal. Scale bars: 50 μm.
Follow JCI:
Copyright © 2019 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts