Efficient generation of NKX6-1+ pancreatic progenitors from multiple human pluripotent stem cell lines

Stem Cell Reports. 2015 Apr 14;4(4):591-604. doi: 10.1016/j.stemcr.2015.02.017. Epub 2015 Apr 2.


Human pluripotent stem cells (hPSCs) represent a renewable source of pancreatic beta cells for both basic research and therapeutic applications. Given this outstanding potential, significant efforts have been made to identify the signaling pathways that regulate pancreatic development in hPSC differentiation cultures. In this study, we demonstrate that the combination of epidermal growth factor (EGF) and nicotinamide signaling induces the generation of NKX6-1(+) progenitors from all hPSC lines tested. Furthermore, we show that the size of the NKX6-1(+) population is regulated by the duration of treatment with retinoic acid, fibroblast growth factor 10 (FGF10), and inhibitors of bone morphogenetic protein (BMP) and hedgehog signaling pathways. When transplanted into NOD scid gamma (NSG) recipients, these progenitors differentiate to give rise to exocrine and endocrine cells, including monohormonal insulin(+) cells. Together, these findings provide an efficient and reproducible strategy for generating highly enriched populations of hPSC-derived beta cell progenitors for studies aimed at further characterizing their developmental potential in vivo and deciphering the pathways that regulate their maturation in vitro.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activins / metabolism
  • Animals
  • Carrier Proteins / metabolism
  • Cell Differentiation*
  • Cell Line
  • Epidermal Growth Factor / metabolism
  • Epidermal Growth Factor / pharmacology
  • Gene Expression Regulation
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Humans
  • Immunophenotyping
  • Islets of Langerhans / cytology
  • Islets of Langerhans / metabolism
  • Mice
  • Models, Biological
  • Niacinamide / pharmacology
  • Organogenesis / drug effects
  • Organogenesis / genetics
  • Pancreas / cytology*
  • Pancreas / metabolism*
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism*
  • Signal Transduction / drug effects


  • Carrier Proteins
  • Homeodomain Proteins
  • NKX6-1 protein, human
  • Activins
  • noggin protein
  • Niacinamide
  • Epidermal Growth Factor