Suppression of Epithelial-to-Mesenchymal Transitioning Enhances Ex Vivo Reprogramming of Human Exocrine Pancreatic Tissue Toward Functional Insulin-Producing β-Like Cells
- Maria João Lima1,
- Kenneth R. Muir1,
- Hilary M. Docherty1,
- Robert Drummond2,
- Neil W.A. McGowan3,
- Shareen Forbes4,
- Yves Heremans5,
- Isabelle Houbracken5,
- James A. Ross2,
- Stuart J. Forbes6,
- Philippe Ravassard7,8,
- Harry Heimberg5,
- John Casey3 and
- Kevin Docherty1⇑
- 1School of Medical Sciences, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen, U.K.
- 2Medical Research Council Centre for Regenerative Medicine, Tissue Injury and Repair Group, University of Edinburgh, Chancellor's Building, Edinburgh, U.K.
- 3Department of Surgery, University of Edinburgh, Edinburgh Royal Infirmary, Edinburgh, U.K.
- 4Endocrinology Unit, University/British Heart Foundation Centre for Cardiovascular Science, Queen’s Medical Research Institute, University of Edinburgh, U.K.
- 5Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium
- 6Medical Research Council Centre for Regenerative Medicine, Scottish Centre for Regenerative Medicine Building, University of Edinburgh, Edinburgh, U.K.
- 7Biotechnology and Biotherapy Laboratory, CNRS UMR 7225, INSERM 975, Paris, France
- 8University Pierre and Marie Curie, Hôpital Pitié Salpêtrière, Paris, France.
- Corresponding author: Kevin Docherty, .
M.J.L. and K.R.M. contributed equally to this study.
Because of the lack of tissue available for islet transplantation, new sources of β-cells have been sought for the treatment of type 1 diabetes. The aim of this study was to determine whether the human exocrine-enriched fraction from the islet isolation procedure could be reprogrammed to provide additional islet tissue for transplantation. The exocrine-enriched cells rapidly dedifferentiated in culture and grew as a mesenchymal monolayer. Genetic lineage tracing confirmed that these mesenchymal cells arose, in part, through a process of epithelial-to-mesenchymal transitioning (EMT). A protocol was developed whereby transduction of these mesenchymal cells with adenoviruses containing Pdx1, Ngn3, MafA, and Pax4 generated a population of cells that were enriched in glucagon-secreting α-like cells. Transdifferentiation or reprogramming toward insulin-secreting β-cells was enhanced, however, when using unpassaged cells in combination with inhibition of EMT by inclusion of Rho-associated kinase (ROCK) and transforming growth factor-β1 inhibitors. Resultant cells were able to secrete insulin in response to glucose and on transplantation were able to normalize blood glucose levels in streptozotocin diabetic NOD/SCID mice. In conclusion, reprogramming of human exocrine-enriched tissue can be best achieved using fresh material under conditions whereby EMT is inhibited, rather than allowing the culture to expand as a mesenchymal monolayer.
This article contains Supplementary Data online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db12-1256/-/DC1.
- Received September 12, 2012.
- Accepted April 14, 2013.
- © 2013 by the American Diabetes Association.
Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.