Abstract
The liver and exocrine pancreas share a common structure, with functioning units (hepatic plates and pancreatic acini) connected to the ductal tree. Here we show that Sox9 is expressed throughout the biliary and pancreatic ductal epithelia, which are connected to the intestinal stem-cell zone. Cre-based lineage tracing showed that adult intestinal cells, hepatocytes and pancreatic acinar cells are supplied physiologically from Sox9-expressing progenitors. Combination of lineage analysis and hepatic injury experiments showed involvement of Sox9-positive precursors in liver regeneration. Embryonic pancreatic Sox9-expressing cells differentiate into all types of mature cells, but their capacity for endocrine differentiation diminishes shortly after birth, when endocrine cells detach from the epithelial lining of the ducts and form the islets of Langerhans. We observed a developmental switch in the hepatic progenitor cell type from Sox9-negative to Sox9-positive progenitors as the biliary tree develops. These results suggest interdependence between the structure and homeostasis of endodermal organs, with Sox9 expression being linked to progenitor status.
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Barker, N. et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449, 1003–1007 (2007).
Sell, S. Alpha-fetoprotein, stem cells and cancer: how study of the production of alpha-fetoprotein during chemical hepatocarcinogenesis led to reaffirmation of the stem cell theory of cancer. Tumour Biol. 29, 161–180 (2008).
Dor, Y., Brown, J., Martinez, O. & Melton, D. Adult pancreatic β cells are formed by self-duplication rather than stem-cell differentiation. Nature 429, 41–46 (2004).
Desai, B.M. et al. Preexisting pancreatic acinar cells contribute to acinar cell, but not islet β cell, regeneration. J. Clin. Invest. 117, 971–977 (2007).
Xu, X. et al. β cells can be generated from endogenous progenitors in injured adult mouse pancreas. Cell 132, 197–207 (2008).
Inada, A. et al. Carbonic anhydrase II-positive pancreatic cells are progenitors for both endocrine and exocrine pancreas after birth. Proc. Natl. Acad. Sci. USA 105, 19915–19919 (2008).
Akiyama, H. Control of chondrogenesis by the transcription factor Sox9. Mod. Rheumatol. 18, 213–219 (2008).
Chaboissier, M.C. et al. Functional analysis of Sox8 and Sox9 during sex determination in the mouse. Development 131, 1891–1901 (2004).
Seymour, P.A. et al. SOX9 is required for maintenance of the pancreatic progenitor cell pool. Proc. Natl. Acad. Sci. USA 104, 1865–1870 (2007).
Antoniou, A. et al. Intrahepatic bile ducts develop according to a new mode of tubulogenesis regulated by the transcription factor SOX9. Gastroenterology 136, 2325–2333 (2009).
Stolt, C.C. et al. The Sox9 transcription factor determines glial fate choice in the developing spinal cord. Genes Dev. 17, 1677–1689 (2003).
Vidal, V.P. et al. Sox9 is essential for outer root sheath differentiation and the formation of the hair stem cell compartment. Curr. Biol. 15, 1340–1351 (2005).
Poché, R.A., Furuta, Y., Chaboissier, M., Schedl, A. & Behringer, R. Sox9 is expressed in mouse multipotent retinal progenitor cells and functions in Müller glial cell development. J. Comp. Neurol. 510, 237–250 (2008).
Foster, J.W. et al. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature 372, 525–530 (1994).
Wagner, T. et al. Autosomal sex reversal and campomelic dysplasia are caused by mutations in and around the SRY-related gene SOX9. Cell 79, 1111–1120 (1994).
Bi, W. et al. Haploinsufficiency of Sox9 results in defective cartilage primordia and premature skeletal mineralization. Proc. Natl. Acad. Sci. USA 98, 6698–6703 (2001).
Mori-Akiyama, Y. et al. SOX9 is required for the differentiation of paneth cells in the intestinal epithelium. Gastroenterology 133, 539–546 (2007).
Nel-Themaat, L. et al. Morphometric analysis of testis cord formation in Sox9-EGFP mice. Dev. Dyn. 238, 1100–1110 (2009).
Kawaguchi, Y., Takaori, K. & Uemoto, S. Genetic lineage tracing, a powerful tool to investigate the embryonic organogenesis and adult organ maintenance of the pancreas. J. Hepatobiliary Pancreat. Sci. published online, doi:10.1007/s00534–010–0307-z (29 July 2010).
Akiyama, H. et al. Osteo-chondroprogenitor cells are derived from Sox9 expressing precursors. Proc. Natl. Acad. Sci. USA 102, 14665–14670 (2005).
Formeister, E.U. et al. Distinct SOX9 levels differentially mark stem/progenitor populations and enteroendocrine cells of the small intestine epithelium. Am. J. Physiol. Gastrointest. Liver Physiol. 296, G1108–G1118 (2009).
van der Flier, L.G. et al. Transcription factor achaete scute-like 2 controls intestinal stem cell fate. Cell 136, 903–912 (2009).
Solar, M. et al. Pancreatic exocrine duct cells give rise to insulin-producing beta cells during embryogenesis but not after birth. Dev. Cell 17, 849–860 (2009).
Raynaud, P., Carpentier, R., Antoniou, A. & Lemaigre, F. Biliary differentiation and bile duct morphogenesis in development and disease. Int. J. Biochem. Cell Biol. published online, doi:10.1016/j.biocel.2009.07.020 (6 September 2009).
Swenson, E.S. et al. Hepatocyte nuclear factor-1 as marker of epithelial phenotype reveals marrow-derived hepatocytes, but not duct cells, after liver injury in mice. Stem Cells 26, 1768–1777 (2008).
Yang, L. et al. Fate-mapping evidence that hepatic stellate cells are epithelial progenitors in adult mouse livers. Stem Cells 26, 2104–2113 (2008).
Petersen, B.E. et al. Bone marrow as a potential source of hepatic oval cells. Science 284, 1168–1170 (1999).
Sackett, S.D. et al. Foxl1 is a marker of bipotential hepatic progenitor cells in mice. Hepatology 49, 920–929 (2009).
Seymour, P.A. et al. A dosage-dependent requirement for Sox9 in pancreatic endocrine cell formation. Dev. Biol. 323, 19–30 (2008).
Kushner, J.A., Weir, G. & Bonner-Weir, S. Ductal origin hypothesis of pancreatic regeneration under attack. Cell Metab. 11, 2–3 (2010).
Zaret, K.S. Genetic programming of liver and pancreas progenitors: lessons for stem-cell differentiation. Nat. Rev. Genet. 9, 329–340 (2008).
Soeda, T. et al. Sox9-expressing precursors are the cellular origin of the cruciate ligament of the knee joint and the limb tendons. Genesis published online, doi:10.1002/dvg.20667 (7 October 2010).
Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat. Genet. 21, 70–71 (1999).
Kawaguchi, Y. et al. The role of the transcriptional regulator Ptf1a in converting intestinal to pancreatic progenitors. Nat. Genet. 32, 128–134 (2002).
Oshima, Y. et al. Isolation of mouse pancreatic ductal progenitor cells expressing CD133 and c-Met by flow cytometric cell sorting. Gastroenterology 132, 720–732 (2007).
Acknowledgements
We thank A. Fukuda and M. Gannon for critical discussions and reading of the manuscript, Y. Ishiura and K. Fukazawa for technical help, C.V.E. Wright for Pdx1 antibody, P. Soriano for Rosa26R mice and the staff of the Institute of Laboratory Animals of Kyoto University for animal care. This work was supported by research grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan and the US National Institutes of Health.
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Y.K. and K.F. designed the study, analyzed the data and prepared the manuscript. K.F. performed the experiments. H.A. and R.R.B. generated mice. T.S. screened mouse lines. J.M.D. cultured embryonic stem cells. E.N., M.H., S.K., T.K., S.H., A.E., M. Koizumi, T.M., M. Kawaguchi, K.T., R.K. and R.D. gave technical support and discussion. T.N. and S.U. supervised the project.
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Furuyama, K., Kawaguchi, Y., Akiyama, H. et al. Continuous cell supply from a Sox9-expressing progenitor zone in adult liver, exocrine pancreas and intestine. Nat Genet 43, 34–41 (2011). https://doi.org/10.1038/ng.722
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DOI: https://doi.org/10.1038/ng.722
