Insulin Production by Human Embryonic Stem Cells
- Suheir Assady1,
- Gila Maor1,
- Michal Amit1,
- Joseph Itskovitz-Eldor12,
- Karl L. Skorecki12 and
- Maty Tzukerman2
- 1Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology, and
- 2Rambam Medical Center, Bat-Galim, Haifa, Israel
Abstract
Type 1 diabetes generally results from autoimmune destruction of pancreatic islet β-cells, with consequent absolute insulin deficiency and complete dependence on exogenous insulin treatment. The relative paucity of donations for pancreas or islet allograft transplantation has prompted the search for alternative sources for β-cell replacement therapy. In the current study, we used pluripotent undifferentiated human embryonic stem (hES) cells as a model system for lineage-specific differentiation. Using hES cells in both adherent and suspension culture conditions, we observed spontaneous in vitro differentiation that included the generation of cells with characteristics of insulin-producing β-cells. Immunohistochemical staining for insulin was observed in a surprisingly high percentage of cells. Secretion of insulin into the medium was observed in a differentiation-dependent manner and was associated with the appearance of other β-cell markers. These findings validate the hES cell model system as a potential basis for enrichment of human β-cells or their precursors, as a possible future source for cell replacement therapy in diabetes.
Footnotes
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Address correspondence and reprint requests to Karl L. Skorecki, Rappaport Research Institute, P.O. Box 9649, Bat-Galim, Haifa 31096, Israel. E-mail: skorecki{at}techunix.technion.ac.il.
Received for publication 16 March 2001 and accepted in revised form 17 May 2001. Posted on the World Wide Web at www.diabetes.org/diabetes on 29 June 2001.
bFGF, basic fibroblast growth factor; dNTP, deoxyribonucleotide; EB, embryoid body; EG, embryonic germ; ES, embryonic stem; GK, glucokinase; hES, human ES; hIns, human insulin; IPF1, insulin promoter factor-1; MEF, mouse embryonic fibroblast; Ngn3, neurogenin-3; Oct4, octamer-binding transcription factor-4; PDX1, pancreatic and duodenal homeobox gene-1; PCR, polymerase chain reaction; RT, reverse transcriptase.














