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.
M Cristina Nostro  1 ,  Farida Sarangi  2 ,  Chaoxing Yang  3 ,  Andrew Holland  4 ,  Andrew G Elefanty  5 ,  Edouard G Stanley  5 ,  Dale L Greiner  3 ,  Gordon Keller  2
Affiliations
  • 1. McEwen Centre for Regenerative Medicine, Toronto, ON M5G 1L7, Canada; Toronto General Research Institute, Department of Experimental Therapeutics, University Health Network, Toronto, ON M5G 1L7, Canada; Department of Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada. Electronic address: [email protected].
  • 2. McEwen Centre for Regenerative Medicine, Toronto, ON M5G 1L7, Canada; Princess Margaret Cancer Centre, University Health Network, Toronto, ON M5G 1L7, Canada.
  • 3. Department of Molecular Medicine and Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • 4. Department of Anatomy and Cell Biology, Monash University, Wellington Road, Clayton, VIC 3800, Australia.
  • 5. Department of Anatomy and Cell Biology, Monash University, Wellington Road, Clayton, VIC 3800, Australia; Murdoch Childrens Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, VIC 3052, Australia.
PMID: 25843049 DOI: 10.1016/j.stemcr.2015.02.017
Abstract

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.