Abstract
Human pluripotent stem cells represent a potentially unlimited source of functional pancreatic endocrine lineage cells. Here we report a highly efficient approach to induce human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells to differentiate into mature insulin-producing cells in a chemical-defined culture system. The differentiated human ES cells obtained by this approach comprised nearly 25% insulin-positive cells as assayed by flow cytometry analysis, which released insulin/C-peptide in response to glucose stimuli in a manner comparable to that of adult human islets. Most of these insulin-producing cells co-expressed mature β cell-specific markers such as NKX6-1 and PDX1, indicating a similar gene expression pattern to adult islet β cells in vivo. In this study, we also demonstrated that EGF facilitates the expansion of PDX1-positive pancreatic progenitors. Moreover, our protocol also succeeded in efficiently inducing human iPS cells to differentiate into insulin-producing cells. Therefore, this work not only provides a new model to study the mechanism of human pancreatic specialization and maturation in vitro, but also enhances the possibility of utilizing patient-specific iPS cells for the treatment of diabetes.
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Acknowledgements
This research was supported by the Ministry of Science and Technology (Grant 2006AA02A113), Science and Technology Plan of Beijing Municipal Government (D07050701350705), Gongjian Project of Beijing Municipal Education Commission, National Basic Research Program of China (973 Program, 2007CB947900, 2009CB941200 and 2009CB941100), National Natural Science Foundation of China (30830061) and a 111 Project to H Deng. We thank Dr Jinning Lou (China-Japan Friendship Hospital) for providing adult human islets and Yizhe Zhang for technical support on real-time PCR. We also thank Yang Zhao, Han Qin, Fangfang Zhu, Zhihua Song, Jiefang You, Yuhua Han and other colleagues in our laboratory for providing technical assistance during the experiments.
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Supplementary information Figure S1
The comparable differentiation efficiency of insulin-producing cells from human ES cell line H1 and H9. (PDF 243 kb)
Supplementary information Figure S2
The pluripotency assay of human iPS cell lines. (PDF 180 kb)
Supplementary information Table S1
Reverse-transcription PCR primers (PDF 62 kb)
Supplementary information Table S2
Quantitive PCR primers (PDF 30 kb)
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Zhang, D., Jiang, W., Liu, M. et al. Highly efficient differentiation of human ES cells and iPS cells into mature pancreatic insulin-producing cells. Cell Res 19, 429–438 (2009). https://doi.org/10.1038/cr.2009.28
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DOI: https://doi.org/10.1038/cr.2009.28
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