In vitro derivation of functional insulin-producing cells from human embryonic stem cells

Cell Res. 2007 Apr;17(4):333-44. doi: 10.1038/cr.2007.28.


The capacity for self-renewal and differentiation of human embryonic stem (ES) cells makes them a potential source for generation of pancreatic beta cells for treating type I diabetes mellitus. Here, we report a newly developed and effective method, carried out in a serum-free system, which induced human ES cells to differentiate into insulin-producing cells. Activin A was used in the initial stage to induce definitive endoderm differentiation from human ES cells, as detected by the expression of the definitive endoderm markers Sox17 and Brachyury. Further, all-trans retinoic acid (RA) was used to promote pancreatic differentiation, as indicated by the expression of the early pancreatic transcription factors pdx1 and hlxb9. After maturation in DMEM/F12 serum-free medium with bFGF and nicotinamide, the differentiated cells expressed islet specific markers such as C-peptide, insulin, glucagon and glut2. The percentage of C-peptide-positive cells exceeded 15%. The secretion of insulin and C-peptide by these cells corresponded to the variations in glucose levels. When transplanted into renal capsules of Streptozotocin (STZ)-treated nude mice, these differentiated human ES cells survived and maintained the expression of beta cell marker genes, including C-peptide, pdx1, glucokinase, nkx6.1, IAPP, pax6 and Tcf1. Thirty percent of the transplanted nude mice exhibited apparent restoration of stable euglycemia; and the corrected phenotype was sustained for more than six weeks. Our new method provides a promising in vitro differentiation model for studying the mechanisms of human pancreas development and illustrates the potential of using human ES cells for the treatment of type I diabetes mellitus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activins / pharmacology
  • Animals
  • Biomarkers / metabolism
  • Cell Differentiation / physiology*
  • Culture Media, Serum-Free
  • DNA-Binding Proteins / metabolism
  • Diabetes Mellitus, Experimental / metabolism
  • Diabetes Mellitus, Experimental / therapy
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Embryonic Stem Cells / transplantation
  • Endoderm / cytology
  • Endoderm / metabolism
  • Fetal Proteins / metabolism
  • High Mobility Group Proteins / metabolism
  • Homeodomain Proteins / metabolism
  • Humans
  • Insulin / biosynthesis*
  • Mice
  • Mice, Nude
  • SOXF Transcription Factors
  • T-Box Domain Proteins / metabolism
  • Trans-Activators / metabolism
  • Transcription Factors / metabolism
  • Tretinoin / pharmacology


  • Biomarkers
  • Culture Media, Serum-Free
  • DNA-Binding Proteins
  • Fetal Proteins
  • High Mobility Group Proteins
  • Homeodomain Proteins
  • Insulin
  • MNX1 protein, human
  • SOX17 protein, human
  • SOXF Transcription Factors
  • T-Box Domain Proteins
  • Trans-Activators
  • Transcription Factors
  • activin A
  • pancreatic and duodenal homeobox 1 protein
  • Activins
  • Tretinoin
  • Brachyury protein