Multipotent stem/progenitor cells in human biliary tree give rise to hepatocytes, cholangiocytes, and pancreatic islets§

Authors

  • Vincenzo Cardinale,

    1. Department of Cell and Molecular Physiology, Biomedical Engineering, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC, USA
    2. Division of Gastroenterology, University Sapienza of Rome, Division of Gastroenterology, Department of Scienze e Biotecnologie Medico-Chirurgiche, Fondazione Eleonora Lorillard Spencer Cenci, Polo Pontino, Rome, Italy, Department of Clinical Medicine, Polo Pontino, Rome, Italy
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    • These authors contributed equally to the study.

  • Yunfang Wang,

    1. Department of Cell and Molecular Physiology, Biomedical Engineering, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC, USA
    Current affiliation:
    1. Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing, PR China, 100850
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  • Guido Carpino,

    1. Department of Health Sciences, University of Rome “Foro Italico”, Rome, Italy
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  • Cai-Bin Cui,

    1. Department of Surgery, UNC School of Medicine, Chapel Hill, NC, USA
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  • Manuela Gatto,

    1. Division of Gastroenterology, University Sapienza of Rome, Division of Gastroenterology, Department of Scienze e Biotecnologie Medico-Chirurgiche, Fondazione Eleonora Lorillard Spencer Cenci, Polo Pontino, Rome, Italy, Department of Clinical Medicine, Polo Pontino, Rome, Italy
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  • Massimo Rossi,

    1. “Paride Stefanini” Department of General Surgery and Organ Transplantation, Sapienza University of Rome, Rome, Italy
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  • Pasquale Bartolomeo Berloco,

    1. “Paride Stefanini” Department of General Surgery and Organ Transplantation, Sapienza University of Rome, Rome, Italy
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  • Alfredo Cantafora,

    1. Division of Gastroenterology, University Sapienza of Rome, Division of Gastroenterology, Department of Scienze e Biotecnologie Medico-Chirurgiche, Fondazione Eleonora Lorillard Spencer Cenci, Polo Pontino, Rome, Italy, Department of Clinical Medicine, Polo Pontino, Rome, Italy
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  • Eliane Wauthier,

    1. Department of Cell and Molecular Physiology, Biomedical Engineering, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC, USA
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  • Mark E. Furth,

    1. Wake Forest Institute of Regenerative Medicine, Winston Salem, NC, USA
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  • Luca Inverardi,

    1. Diabetes Research Institute, University of Miami, Miami, FL, USA
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  • Juan Dominguez-Bendala,

    1. Diabetes Research Institute, University of Miami, Miami, FL, USA
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  • Camillo Ricordi,

    1. Diabetes Research Institute, University of Miami, Miami, FL, USA
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  • David Gerber,

    1. Department of Surgery, UNC School of Medicine, Chapel Hill, NC, USA
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  • Eugenio Gaudio,

    1. Department of Health Sciences, University of Rome “Foro Italico”, Rome, Italy
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    • Coequal senior authors.

  • Domenico Alvaro,

    Corresponding author
    1. Division of Gastroenterology, University Sapienza of Rome, Division of Gastroenterology, Department of Scienze e Biotecnologie Medico-Chirurgiche, Fondazione Eleonora Lorillard Spencer Cenci, Polo Pontino, Rome, Italy, Department of Clinical Medicine, Polo Pontino, Rome, Italy
    • Division of Gastroenterology, University Sapienza of Rome, Division of Gastroenterology, Department of Scienze e Biotecnologie Medico-Chirurgiche, Fondazione Eleonora Lorillard Spencer Cenci, Polo Pontino, Rome, Italy
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    • Coequal senior authors.

    • fax: 011 39 06 4453319

  • Lola Reid

    Corresponding author
    1. Department of Cell and Molecular Physiology, Biomedical Engineering, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC, USA
    2. Wake Forest Institute of Regenerative Medicine, Winston Salem, NC, USA
    • Department of Cell and Molecular Physiology, RM 34 Glaxo, UNC School of Medicine, Chapel Hill, NC 27599
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    • These authors contributed equally to the study.

    • fax: 919-966-6112


  • Potential conflict of interest: Dr. Reid consults, received grants from, and holds intellectuals property rights for Vesta and GigaCyte. She also received grants from and holds intellectual property rights for Vertex.

  • Support: (UNC) Funding derived from a grant from the North Carolina Biotechnology Center (NCBC), GigaCyte Biotech (Branford, CT), Vesta Therapeutics (Bethesda, MD), and from NIH grants (AA014243, IP30-DK065933), NIDDK Grant (DK34987), and an NCI grant (CA016086); (Sapienza University) Dr. Cardinale received salary support from a scholarship from Sapienza University of Rome for the studies that he did at UNC. D. Alvaro and V. Cardinale were supported by FIRB grant no. RBAP10Z7FS_004; D. Alvaro, V. Cardinale, E. Gaudio, and G. Carpino were supported by a grant from Agenzia Regionale Del Lazio Per I Trapianti E Le Patologie Connesse; E. Gaudio was supported by MIUR grants: PRIN#2007, prot. 2007HPT7BA_001 and Federate Athenaeum funds from the University Sapienza of Rome; (Diabetes Research Institute) The studies were funded by grants from NIH, the Juvenile Diabetes Research Foundation, ADA, and the Diabetes Research Institute Foundation.

  • §

    Patent: A patent on the biliary tree stem cells was filed in November, 2009 and is jointly owned by UNC in Chapel Hill, NC, and Sapienza University in Rome, Italy.

Abstract

Multipotent stem/progenitors are present in peribiliary glands of extrahepatic biliary trees from humans of all ages and in high numbers in hepato-pancreatic common duct, cystic duct, and hilum. They express endodermal transcription factors (e.g., Sox9, SOX17, FOXA2, PDX1, HES1, NGN3, PROX1) intranuclearly, stem/progenitor surface markers (EpCAM, NCAM, CD133, CXCR4), and sometimes weakly adult liver, bile duct, and pancreatic genes (albumin, cystic fibrosis transmembrane conductance regulator [CFTR], and insulin). They clonogenically expand on plastic and in serum-free medium, tailored for endodermal progenitors, remaining phenotypically stable as undifferentiated cells for months with a cell division initially every ≈36 hours and slowing to one every 2-3 days. Transfer into distinct culture conditions, each comprised of a specific mix of hormones and matrix components, yields either cords of hepatocytes (express albumin, CYP3A4, and transferrin), branching ducts of cholangiocytes (expressing anion exchanger-2-AE2 and CFTR), or regulatable C-peptide secreting neoislet-like clusters (expressing glucagon, insulin) and accompanied by changes in gene expression correlating with the adult fate. Transplantation into quiescent livers of immunocompromised mice results in functional human hepatocytes and cholangiocytes, whereas if into fat pads of streptozocin-induced diabetic mice, results in functional islets secreting glucose-regulatable human C-peptide. Conclusion: The phenotypes and availability from all age donors suggest that these stem/progenitors have considerable potential for regenerative therapies of liver, bile duct, and pancreatic diseases including diabetes. (HEPATOLOGY2011;)

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