title: Evidence for multipotent endodermal stem/progenitor cell populations in human gallbladder.
abstract: OBJECTIVE
  Multipotent stem/progenitor cells are found in peribiliary glands throughout human biliary trees and are able to generate mature cells of hepato-biliary and pancreatic endocrine lineages. The presence of endodermal stem/progenitors in human gallbladder was explored.
  
  
  METHODS
  Gallbladders were obtained from organ donors and laparoscopic surgery for symptomatic cholelithiasis. Tissues or isolated cells were characterized by immunohistochemistry and flow cytometry. EpCAM+ (Epithelial Cell Adhesion Molecule) cells were immunoselected by magnetic microbeads, plated onto plastic in self-replication conditions and subsequently transferred to distinct serum-free, hormonally defined media tailored for differentiation to specific adult fates. In vivo studies were conducted in an experimental model of liver cirrhosis.
  
  
  RESULTS
  The gallbladder does not have peribiliary glands, but it has stem/progenitors organized instead in mucosal crypts. Most of these can be isolated by immune-selection for EpCAM. Approximately 10% of EpCAM+ cells in situ and of immunoselected EpCAM+ cells co-expressed multiple pluripotency genes and various stem cell markers; other EpCAM+ cells qualified as progenitors. Single EpCAM+ cells demonstrated clonogenic expansion ex vivo with maintenance of stemness in self-replication conditions. Freshly isolated or cultured EpCAM+ cells could be differentiated to multiple, distinct adult fates: cords of albumin-secreting hepatocytes, branching ducts of secretin receptor+ cholangiocytes, or glucose-responsive, insulin/glucagon-secreting neoislets. EpCAM+ cells transplanted in vivo in immune-compromised hosts gave rise to human albumin-producing hepatocytes and to human Cytokeratin7+ cholangiocytes occurring in higher numbers when transplanted in cirrhotic mice.
  
  
  CONCLUSIONS
  Human gallbladders contain easily isolatable cells with phenotypic and biological properties of multipotent, endodermal stem cells.
authors: Department of Movement, Human and Health Sciences, Division of Health Sciences, University of Rome "Foro Italico", Rome, Italy.
authors: Guido
authors: G
authors: Carpino
authors: Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy.
authors: Vincenzo
authors: V
authors: Cardinale
authors: Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy.
authors: Raffaele
authors: R
authors: Gentile
authors: Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy; Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy.
authors: Paolo
authors: P
authors: Onori
authors: Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy.
authors: Rossella
authors: R
authors: Semeraro
authors: Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy; Eleonora Lorillard Spencer-Cenci Foundation, Rome, Italy.
authors: Antonio
authors: A
authors: Franchitto
authors: Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC 27599, United States.
authors: Yunfang
authors: Y
authors: Wang
authors: Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.
authors: Daniela
authors: D
authors: Bosco
authors: Surgical-Medical Department for Digestive Diseases, Sapienza University of Rome, Rome, Italy.
authors: Angelo
authors: A
authors: Iossa
authors: Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.
authors: Chiara
authors: C
authors: Napoletano
authors: Division of Gastroenterology, Sapienza University of Rome, Rome, Italy.
authors: Alfredo
authors: A
authors: Cantafora
authors: Gastroenterology Unit, Department of Clinical and Experimental Medicine, Federico II University of Naples, Italy.
authors: Giuseppe
authors: G
authors: D'Argenio
authors: Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.
authors: Marianna
authors: M
authors: Nuti
authors: Gastroenterology Unit, Department of Clinical and Experimental Medicine, Federico II University of Naples, Italy.
authors: Nicola
authors: N
authors: Caporaso
authors: Department of General Surgery and Organ Transplantation, Sapienza University of Rome, Rome, Italy.
authors: Pasquale
authors: P
authors: Berloco
authors: Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy.
authors: Rosanna
authors: R
authors: Venere
authors: Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC 27599, United States.
authors: Tsunekazu
authors: T
authors: Oikawa
authors: Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC 27599, United States.
authors: Lola
authors: L
authors: Reid
authors: Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy; Eleonora Lorillard Spencer-Cenci Foundation, Rome, Italy.
authors: Domenico
authors: D
authors: Alvaro
authors: Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy. Electronic address: eugenio.gaudio@uniroma1.it.
authors: Eugenio
authors: E
authors: Gaudio
mesh_terms: D000818:Animals; D001659:Biliary Tract / Q000166:cytology; D002454:Cell Differentiation; D002769:Cholelithiasis / Q000473:pathology / Q000601:surgery; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000166:cytology; D005704:Gallbladder / Q000166:cytology*; D019073:HT29 Cells; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D018189:Immunomagnetic Separation; D007515:Islets of Langerhans / Q000166:cytology; D008106:Liver Cirrhosis, Experimental / Q000473:pathology / Q000628:therapy*; D008115:Liver Regeneration; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology*; D061251:Primary Cell Culture; D055153:Stem Cell Niche; D014019:Tissue Donors
keywords: Epithelial differentiation; Gallbladder; Liver regeneration; Pancreatic islet cells; Stem cells