HES-1 is involved in adaptation of adult human beta-cells to proliferation in vitro

Diabetes. 2008 Sep;57(9):2413-20. doi: 10.2337/db07-1323. Epub 2008 Jul 3.


Objective: In vitro expansion of beta-cells from adult human islets could solve the tissue shortage for cell replacement therapy of diabetes. Culture of human islet cells typically results in <16 cell doublings and loss of insulin expression. Using cell lineage tracing, we demonstrated that the expanded cell population included cells derived from beta-cells. Understanding the molecular mechanisms involved in beta-cell fate in vitro is crucial for optimizing expansion and redifferentiation of these cells. In the developing pancreas, important cell-fate decisions are regulated by NOTCH receptors, which signal through the hairy and enhancer of split (HES)-1 transcriptional regulator. Here, we investigated the role of the NOTCH signaling pathway in beta-cell dedifferentiation and proliferation in vitro.

Research design and methods: Isolated human islets were dissociated into single cells. beta-Cells were genetically labeled using a Cre-lox system delivered by lentiviruses. Cells were analyzed for changes in expression of components of the NOTCH pathway during the initial weeks in culture. HES-1 expression was inhibited by a small hairpin RNA (shRNA), and the effects on beta-cell phenotype were analyzed.

Results: Human beta-cell dedifferentiation and entrance into the cell cycle in vitro correlated with activation of the NOTCH pathway and downregulation of the cell cycle inhibitor p57. Inhibition of HES-1 expression using shRNA resulted in significantly reduced beta-cell replication and dedifferentiation.

Conclusions: These findings demonstrate that the NOTCH pathway is involved in determining beta-cell fate in vitro and suggest possible molecular targets for induction of beta-cell redifferentiation following in vitro expansion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Basic Helix-Loop-Helix Transcription Factors / genetics*
  • Basic Helix-Loop-Helix Transcription Factors / metabolism*
  • Cell Culture Techniques
  • Cell Differentiation / physiology
  • Cell Division / physiology
  • Cell Lineage / physiology
  • Cells, Cultured
  • Cyclin-Dependent Kinase Inhibitor p27 / metabolism
  • Homeodomain Proteins / genetics*
  • Homeodomain Proteins / metabolism*
  • Humans
  • Insulin-Secreting Cells / cytology*
  • Insulin-Secreting Cells / physiology*
  • RNA, Small Interfering
  • Receptors, Notch / metabolism
  • Signal Transduction / physiology*
  • Transcription Factor HES-1
  • Up-Regulation / physiology


  • Basic Helix-Loop-Helix Transcription Factors
  • Homeodomain Proteins
  • RNA, Small Interfering
  • Receptors, Notch
  • Transcription Factor HES-1
  • Cyclin-Dependent Kinase Inhibitor p27
  • HES1 protein, human