Directed Differentiation of Embryonic Stem Cells Allows Exploration of Novel Transcription Factor Genes for Pancreas Development

被引:0
作者
Jing Sui
Munish Mehta
Bingyin Shi
Grant Morahan
Fang-Xu Jiang
机构
[1] University of Western Australia,Centre for Diabetes Research, The Western Australian Institute for Medical Research, Centre for Medical Research
[2] First Affiliated Hospital of Medical College of Xi’an Jiaotong University,Department of Endocrinology
来源
Stem Cell Reviews and Reports | 2012年 / 8卷
关键词
Islet Cell; Leukemia Inhibitory Factor; DAPT; Cyclopamine; Definitive Endoderm;
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摘要
Embryonic stem cells (ESCs) have been promised as a renewable source for regenerative medicine, including providing a replacement therapy in type 1 diabetes. However, they have not yet been differentiated into functional insulin-secreting β cells. This is due partially to the knowledge gap regarding the transcription factors (TFs) required for pancreas development. We hypothesize that, if directed differentiation in vitro recapitulates the developmental process in vivo, ESCs provide a powerful model to discover novel pancreatic TF genes. Guided by knowledge of their normal development and using RT-PCR and immunochemical analyses, we have established protocols for directed differentiation of mouse ESCs into pancreatic progenitors. Microarray analyses of these differentiating ESC cells at days 0, 4, 8 and 15 confirmed their sequential differentiation. By day 15, we found up-regulation of a group of pancreatic progenitor marker genes including Pdx1, Ptf1a, Nkx6.1, Pax4 and Pax6. Consistently, Pdx1-immunoreactive cells were detected on day 15. Most of these Pdx1+ cells also expressed Nkx6.1. Bioinformatic analyses of sequential datasets allowed identification of over 20 novel TF genes potentially important for pancreas development. The dynamic expression of representative known and novel genes was confirmed by quantitative real time RT-PCR analysis. This strategy may be modified to study novel regulatory molecules for development of other tissue and organ systems.
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页码:803 / 812
页数:9
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[1]  
D’Amour KA(2006)Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells Nature Biotechnology 24 1392-1401
[2]  
Bang AG(2008)Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo Nature Biotechnology 26 443-452
[3]  
Eliazer S(2009)A small molecule that directs differentiation of human ESCs into the pancreatic lineage Nature Chemical Biology 5 258-265
[4]  
Kroon E(2009)Small molecules efficiently direct endodermal differentiation of mouse and human embryonic stem cells Cell Stem Cell 4 348-358
[5]  
Martinson LA(2002)Signal transduction and the control of gene expression Science 295 813-818
[6]  
Kadoya K(2002)Extracellular signals and pancreatic beta-cell development: A brief review Molecular Medicine 8 763-770
[7]  
Chen S(1972)An ultrastructural analysis of the developing embryonic pancreas Developmental Biology 29 436-467
[8]  
Borowiak M(2002)Direct evidence for the pancreatic lineage: NGN3+ cells are islet progenitors and are distinct from duct progenitors Development 129 2447-2457
[9]  
Fox JL(2000)The homeodomain of PDX-1 mediates multiple protein-protein interactions in the formation of a transcriptional activation complex on the insulin promoter Molecular and Cellular Biology 20 900-911
[10]  
Borowiak M(2000)neurogenin3 is required for the development of the four endocrine cell lineages of the pancreas Proceedings of the National Academy of Sciences of the United States of America 97 1607-1611