Efficient Generation of Glucose-Responsive Beta Cells from Isolated GP2+ Human Pancreatic Progenitors

Research output: Contribution to journalArticle

Abstract

Stem cell-based therapy for type 1 diabetes would benefit from implementation of a cell purification step at the pancreatic endoderm stage. This would increase the safety of the final cell product, allow the establishment of an intermediate-stage stem cell bank, and provide a means for upscaling β cell manufacturing. Comparative gene expression analysis revealed glycoprotein 2 (GP2) as a specific cell surface marker for isolating pancreatic endoderm cells (PECs) from differentiated hESCs and human fetal pancreas. Isolated GP2+ PECs efficiently differentiated into glucose responsive insulin-producing cells in vitro. We found that in vitro PEC proliferation declines due to enhanced expression of the cyclin-dependent kinase (CDK) inhibitors CDKN1A and CDKN2A. However, we identified a time window when reducing CDKN1A or CDKN2A expression increased proliferation and yield of GP2+ PECs. Altogether, our results contribute tools and concepts toward the isolation and use of PECs as a source for the safe production of hPSC-derived β cells.

Details

Authors
  • Jacqueline Ameri
  • Rehannah Borup
  • Christy Prawiro
  • Cyrille Ramond
  • Karen A. Schachter
  • Raphael Scharfmann
  • Henrik Semb
Organisations
External organisations
  • University of Copenhagen
  • Copenhagen University Hospital
  • Paris Descartes University
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Cell and Molecular Biology

Keywords

  • CDKN1A, CDKN2A, cell surface marker, cell-cycle regulators, differentiation, GP2, human embryonic stem cells, insulin-producing beta cells, pancreatic progenitors, type 1 diabetes
Original languageEnglish
Pages (from-to)36-49
Number of pages14
JournalCell Reports
Volume19
Issue number1
Publication statusPublished - 2017 Apr 4
Publication categoryResearch
Peer-reviewedYes