• Open Access

Biliary tree stem cells, precursors to pancreatic committed progenitors: Evidence for possible life-long pancreatic organogenesis

Authors

  • Yunfang Wang,

    1. Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, Lineberger Cancer Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA
    Search for more papers by this author
  • Giacomo Lanzoni,

    1. Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, Florida, USA
    2. Department of Histology, Embryology and Applied Biology, University of Bologna, Bologna, Italy
    Search for more papers by this author
  • Guido Carpino,

    1. Department of Health Sciences, University of Rome “ForoItalico”, Rome, Italy
    Search for more papers by this author
  • Cai-Bin Cui,

    1. Department of Surgery, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA
    Search for more papers by this author
  • Juan Dominguez-Bendala,

    1. Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, Florida, USA
    Search for more papers by this author
  • Eliane Wauthier,

    1. Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, Lineberger Cancer Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA
    Search for more papers by this author
  • Vincenzo Cardinale,

    1. Department of Medico-Surgical Sciences and Biotechnologies, Sapienza University, Rome, Italy
    Search for more papers by this author
  • Tsunekazu Oikawa,

    1. Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, Lineberger Cancer Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA
    Search for more papers by this author
  • Antonello Pileggi,

    1. Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, Florida, USA
    Search for more papers by this author
  • David Gerber,

    1. Department of Surgery, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA
    Search for more papers by this author
  • Mark E. Furth,

    1. Innovation Wake Forest Innovations, Wake Forest Baptist Medical Center, Winston Salem, North Carolina, USA
    Search for more papers by this author
  • Domenico Alvaro,

    1. Department of Medico-Surgical Sciences and Biotechnologies, Sapienza University, Rome, Italy
    Search for more papers by this author
  • Eugenio Gaudio,

    1. Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University, Rome, Italy
    Search for more papers by this author
  • Luca Inverardi,

    1. Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, Florida, USA
    Search for more papers by this author
  • Lola M. Reid

    Corresponding author
    1. Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, Lineberger Cancer Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA
    • Correspondence: Lola M. Reid, Department of Cell Biology and Physiology and Program in Molecular Biology and Biotechnology, CB# 7038, UNC School of Medicine, Glaxo Research Building, Rm. 34, Chapel Hill, North Carolina 27599-7038, USA. Telephone: 919-966-0346; Fax: 919-966-6112; e-mail: Lola.M.Reid@gmail.com

    Search for more papers by this author

  • Author contributions: Y.W.: original ideas, conception and design of the project, acquisition and assembly of data and data analyses, interpretation, manuscript writing and editing. Efforts with senior authors for final approval of manuscript; G.L., G.C, and C.C.: key investigators (co-equal second authors) in acquisition and assembly of data and data analyses; interpretation; manuscript writing and editing; V.C. J.B., and T.O.: acquisition and assembly of data and figures and data analyses; interpretation; manuscript writing and editing. E.W. and A.P., acquisition and assembly of data and figures and data analyses; D.G.: manuscript writing and editing; M.F. (senior author): literature assembly and analyses; experimental designs; data analyses; manuscript writing and editing; D.A., E.G. L.I. (senior authors): conception and design, assembly of data, data analyses and interpretation, manuscript editing, final approval of manuscript and financial support; L.R. (senior and corresponding author): original ideas; conception and design of the project; data analyses and interpretation, initial preparation of manuscript and all stages of manuscript editing, final approval of manuscript and financial support.

Abstract

Peribiliary glands (PBGs) in bile duct walls, and pancreatic duct glands (PDGs) associated with pancreatic ducts, in humans of all ages, contain a continuous, ramifying network of cells in overlapping maturational lineages. We show that proximal (PBGs)-to-distal (PDGs) maturational lineages start near the duodenum with cells expressing markers of pluripotency (NANOG, OCT4, and SOX2), proliferation (Ki67), self-replication (SALL4), and early hepato-pancreatic commitment (SOX9, SOX17, PDX1, and LGR5), transitioning to PDG cells with no expression of pluripotency or self-replication markers, maintenance of pancreatic genes (PDX1), and expression of markers of pancreatic endocrine maturation (NGN3, MUC6, and insulin). Radial-axis lineages start in PBGs near the ducts' fibromuscular layers with stem cells and end at the ducts' lumens with cells devoid of stem cell traits and positive for pancreatic endocrine genes. Biliary tree-derived cells behaved as stem cells in culture under expansion conditions, culture plastic and serum-free Kubota's Medium, proliferating for months as undifferentiated cells, whereas pancreas-derived cells underwent only approximately 8–10 divisions, then partially differentiated towards an islet fate. Biliary tree-derived cells proved precursors of pancreas' committed progenitors. Both could be driven by three-dimensional conditions, islet-derived matrix components and a serum-free, hormonally defined medium for an islet fate (HDM-P), to form spheroids with ultrastructural, electrophysiological and functional characteristics of neoislets, including glucose regulatability. Implantation of these neoislets into epididymal fat pads of immunocompromised mice, chemically rendered diabetic, resulted in secretion of human C-peptide, regulatable by glucose, and able to alleviate hyperglycemia in hosts. The biliary tree-derived stem cells and their connections to pancreatic committed progenitors constitute a biological framework for life-long pancreatic organogenesis. Stem Cells 2013;31:1966-1979

Ancillary