Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo

Abstract

Development of a cell therapy for diabetes would be greatly aided by a renewable supply of human β-cells. Here we show that pancreatic endoderm derived from human embryonic stem (hES) cells efficiently generates glucose-responsive endocrine cells after implantation into mice. Upon glucose stimulation of the implanted mice, human insulin and C-peptide are detected in sera at levels similar to those of mice transplanted with 3,000 human islets. Moreover, the insulin-expressing cells generated after engraftment exhibit many properties of functional β-cells, including expression of critical β-cell transcription factors, appropriate processing of proinsulin and the presence of mature endocrine secretory granules. Finally, in a test of therapeutic potential, we demonstrate that implantation of hES cell–derived pancreatic endoderm protects against streptozotocin-induced hyperglycemia. Together, these data provide definitive evidence that hES cells are competent to generate glucose-responsive, insulin-secreting cells.

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Figure 1: hES cell differentiation to pancreatic endoderm.
Figure 2: Human C-peptide and insulin in sera of implanted mice.
Figure 3: Morphological and immunofluorescence analyses of graft samples 78 d post implant (Supplementary Table 1, mouse no. 14).
Figure 4: Immunofluorescence analysis of islet amyloid polypeptide (IAPP) expression in hES cell–derived pancreatic endocrine cells.
Figure 5: Ultrastructural and immunofluorescence analyses of grafts.
Figure 6: Analyses of STZ-treated mice.

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Acknowledgements

We thank Ole Madsen (Hagedorn Research Institute), Christopher Wright (Vanderbilt University), Roland Stein (Vanderbilt University) and Patrick Collombat (Max-Planck Institute) for antibody reagents; Michael McCaffery at John Hopkins University for performing the TEM analyses. The CyT203 and CyT49 cell lines were derived with partial funding from the Juvenile Diabetes Research Foundation.

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The authors are full-time employees at Novocell Inc., a biotechnology company.

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Supplementary Figures 1–4, Tables 1–4 (PDF 9234 kb)

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Kroon, E., Martinson, L., Kadoya, K. et al. Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo. Nat Biotechnol 26, 443–452 (2008). https://doi.org/10.1038/nbt1393

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