Abstract
Stepwise differentiation from embryonic stem cells (ESCs) to functional insulin-secreting beta cells will identify key steps in beta-cell development and may yet prove useful for transplantation therapy for diabetics. An essential step in this schema is the generation of pancreatic progenitors—cells that express Pdx1 and produce all the cell types of the pancreas. High-content chemical screening identified a small molecule, (−)-indolactam V, that induces differentiation of a substantial number of Pdx1-expressing cells from human ESCs. The Pdx1-expressing cells express other pancreatic markers and contribute to endocrine, exocrine and duct cells, in vitro and in vivo. Further analyses showed that (−)-indolactam V works specifically at one stage of pancreatic development, inducing pancreatic progenitors from definitive endoderm. This study describes a chemical screening platform to investigate human ESC differentiation and demonstrates the generation of a cell population that is a key milepost on the path to making beta cells.
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Acknowledgements
D.M. and S.L.S. are investigators of the Howard Hughes Medical Institute. S.C. is supported by the Howard Hughes Medical Institute and Juvenile Diabetes Research Foundation. R.M. is supported by the Howard Hughes Medical Institute. L.F.P. is supported by the American Gastroenterological Association and the National Pancreas Foundation. The authors thank D. Huangfu for thoughtful discussion, H. Ngo for assistance with the high-content screen, S. Huang for help with NMR analysis and the Beta Cell Biology Consortium for providing antibodies.
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S.C., D.M. and S.L.S. designed the experiments, analyzed data and wrote the manuscript; S.C., M.B., R.M. and K.O. performed the experiments; J.L.F., L.F.P., S.L.S. and L.L.R. provided the chemical library; and S.C., L.D., K.L. and L.L.R. performed the high-content screen.
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The authors have filed a patent on the ILV molecule described in the paper.
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Chen, S., Borowiak, M., Fox, J. et al. A small molecule that directs differentiation of human ESCs into the pancreatic lineage. Nat Chem Biol 5, 258–265 (2009). https://doi.org/10.1038/nchembio.154
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DOI: https://doi.org/10.1038/nchembio.154
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