Abstract
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.
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Acknowledgements
This research was supported by the Ministry of Science and Technology Grant (2001CB510106), Science and Technology Plan of Beijing Municipal Government (H020220050290), National Natural Science Foundation of China Awards for Outstanding Young Scientists (30125022) and for Creative Research Groups (30421004), and Bill & Melinda Gates Foundation Grant (37871) to H Deng. We thank Dr C Wright for kindly providing us with the pdx1 antibody. We thank Drs Matt Stremlau, Tung-Tien Sun, Johnny He, and Hui Zhang for critical reading of the manuscript. We thank Yizhe Zhang for technical support on real-time PCR. We also thank Donghui Zhang, Xulei Sun, Yushan Guo, Zan Tong, Jun Cai, Han Qin, Chengyan Wang, Pengbo Zhang, Chunbo Teng, Jiefang You, Yan Shen, Wei Wei and other colleagues in our laboratory for technical assistance and advice during experiments.
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Jiang, W., Shi, Y., Zhao, D. et al. In vitro derivation of functional insulin-producing cells from human embryonic stem cells. Cell Res 17, 333–344 (2007). https://doi.org/10.1038/cr.2007.28
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DOI: https://doi.org/10.1038/cr.2007.28
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